Amino- or ammonium-containing sulfonic acid, phosphonic acid and carboxylic acid derivatives and their medical use

ABSTRACT

The present invention relates to amino- or ammonium-containing sulfonic acid, phosphonic acid and carboxylic acid derivatives, in particular the compounds of formula 1, 2, 3, 4, 5 or 6 and their medical use, including their use in treatment, prevention or amelioration of an inflammatory, autoimmune and/or allergic disorder.

This application is a divisional of U.S. patent application Ser. No.14/403,167, which was filed Apr. 27, 2015, which is a 371 ofinternational application PCT/ EP2012/059812 filed May 25, 2012, whichclaims priority to and the benefit of EP 11167752.2 filed May 26, 2011,the entire contents of all of which are incorporated herein by reference

The present invention relates to amino- or ammonium-containing sulfonicacid, phosphonic acid and carboxylic acid derivatives, in particular thecompounds of formula 1, 2, 3, 4, 5 or 6, and their medical use,including their use in the treatment, prevention or amelioration of aninflammatory, autoimmune and/or allergic disorder.

Without being bound by theory, the compounds provided herein areconsidered to exert their pharmacological activity through inhibition ofthe phosphoinosftide 3-kinase (PI3K)/Akt kinase pathway. Theserine/threonine protein kinase Akt (also known as Protein Kinase B) isa key mediator of signal transduction. Akt is activated by numerousreceptors, including those of growth factors, cytokines, hormones andinsulin as well as by the attachment of cells to the extracellularmatrix. Once activated, the plasma membrane receptors stimulate theactivity of PI3K to generate phosphatidylinositol-3,4,5-trisphosphate(PIP3), a lipid second messenger essential for the translocation of Akt,which contains a PlP3-binding pleckstrin homology (PH)-domain, from thecytoplasm to the plasma membrane (Franke et al., Cell 81 :727-736,(1995)). Once recruited to the membrane, it is phosphoryiated andactivated by other kinases (Hemmings, Science 275:628-630 (1997);Hemmlngs, Science 276:534 (1997); Downward, Science 279:673-674 (1998);Alessi et al., EMBO J. 15:6541- 6551 (1996)), such as PDK1 and mTORC2.

Akt in turn is responsible for regulating the function of many cellularproteins involved in processes such as transcription and apoptosis(programmed cell death), angiogenesis, cell motility and glucosemetabolism (Kulik et al., Mol Cell Biol. 17:1595-1606 (1997); Franke etal., Cell 88:435-437 (1997); Kaufmann-Zeh et al., Nature 385:544-548(1997) Hemmings, Science 275:628-630 (1997); Dudek et al., Science275:661-665 (1997)).

These findings indicate that Akt may be a drug target for the treatmentof inflammation, autoimmune diseases and allergy. Accordingly, thecompounds provided herein, in particular the compounds of formula 1, 2,3, 4, 5 or 6, are useful in the treatment, prevention or amelioration ofsuch illnesses.

Broad-acting immunomodulatory drugs such as corticosteroids, calcineurininhibitors and cyclosporin are highly effective and have been used forthe therapy of allergic and cellular inflammatory diseases, includingautoimmune diseases, for many years. They are potent in suppressing bothTh1 and Th2 driven processes, yet they suffer from undesirableside-effects, which limit their therapeutic window. Corticosteroidsregulate the expression of numerous genes and, consequently, their useis limited by severe adverse effects. Typical serious adverse effects ofshort-term corticosteroids use are disturbances in water and saltretention, lipid metabolism, skin thinning and changes in behaviour.More serious adverse effects associated with long-term systemic exposureto corticosteroids include increased appetite and weight gain, depositsof fat in chest, face, upper back, and stomach, water and salt retentionleading to swelling and edema, high blood pressure, diabetes, slowedhealing of wounds, osteoporosis, cataracts, acne, muscle weakness,thinning of the skin, increased susceptibility to infection, stomachulcers, increased sweating, mood swings, psychological problems such asdepression, adrenal suppression and crisis.

More specifically targeted therapeutics, such as biologies, e.g.,antibodies against certain cytokines or their receptors, inhibit asingle protein target and are effective in certain situations, but onlyaddress one of the targets in a highly redundant inflammatory cascadeand are hence often used in combination therapy, as effective resolutionof inflammatory diseases requires several targets to be addressedsimultaneously.

There is a high unmet medical need for new drugs that curb theunderlying disease processes. For instance, in rheumatoid arthritis (RA)such disease-modifying antirheumatic drugs (DMARDs) can slow downprogressive joint destruction reducing long-term disease severity. Thisprovides both therapeutic and economic advantages by shortening thetherapeutic period and reducing the dose of concomitant medications.

Many chronic inflammatory diseases, including autoimmune diseases suchas rheumatoid arthritis (RA), are associated with deregulatedintracellular signal transduction pathways, including thephosphoinositide 3-kinase (PI3K)/Akt kinase pathway, and the resultantpathogenic interactions between immune and connective tissue stromalcells lead to changes in cell activation, proliferation, migratorycapacity, and cell survival that contribute to inflammation (Tas et aL,Curr Pharm Des. 11:581-611 (2005)). For example, abnormal functioning,differentiation and/or activation of T-ceils, B-cells and myeloid cellshave been documented in various autoimmune diseases, includingrheumatoid arthritis (RA), diabetes mellitus, lupus and multiplesclerosis and studies have detailed anomalous activation of the Aktsignalling axis in the context of systemic autoimmunity (Wu et al.,Disord Drug Targets. 9:145-50 (2009)).

Akt is an important signal transduction pathway mediating the delay ofneutrophil apoplosis by inflammatory mediators, during neutrophilactivation during inflammation (Rane and Klein, Front Biosci. 14:2400-12(2009)) and control over neutrophil and macrophage migration andapoptosis is a key factor in the pathogenesis of the majority of chronicinflammatory diseases.

RA is a chronic inflammatory disease, which results in inflammation ofthe synovial lining and destruction of the adjacent bone and cartilage.Synovial macrophages, fibroblasts and lymphocytes are critical to thepathogenesis of this disease, in which apoptosis plays divergent roles.Signaling pathways, such as PI3K/Akt, are highly activated in the RAjoint, contributing to the expression of genes that cause inflammationand destruction and expression of a variety of anti-apoptotic molecules.Induction of apoptosis of macrophages, synovial fibroblasts orlymphocytes, through inhibition of the expression of anti-apoptoticmolecules, could be therapeutically beneficial in RA (Liu and Pope, CurrOpin Pharmacol. 3:317-22 (2003)). Furthermore, results suggest thatsignal transduction pathways dependent on PI3K/Akt are involved in theoverproduction of the key inflammatory cytokine IL-17 in patients withrheumatoid arthritis (Kim et al., Arthritis Res Ther. 7:R139-148(2005)).

Akt is closely associated with key membrane-bound receptors andrepresents a convergent integration point for multiple stimuliimplicated in COPD pathogenesis. Akt is also implicated in the systemicmanifestations of COPD such as skeletal muscle wasting and metabolicdisturbances. As such, Akt represents a particularly attractivetherapeutic target for the treatment of COPD (Bozinovski et al, fnt JChron Obstruct Pulmon Dis. 1:31-38 (2006)).

The compounds provided herein are positioned to be disease-modifyingdrugs. The compounds have potential for application in a wide variety ofchronic inflammatory indications and in combination with a favorabletolerability, the products can be expected to gain adoption by asignificant number of patients suffering from the severe side effects ofcurrent treatments. Furthermore, the compounds provided herein will besuitable not only for monotherapy but also in combination with existingtherapies, which address specific disease targets but are not sufficientto resolve the disease alone.

Activation of the PI3K/Akt pathway is essential for insulin-inducedglucose metabolism, including translocation of GLUT4 transported to theceil surface, glucose uptake, glycogen synthesis, suppression of glucoseoutput and triglyceride synthesis as well as insulin-induced mitogenesis(Asano et al., Bioi Pharm Bull. 30:1610-6 (2007)). Hence, inhibitors ofPI3K/Akt signalling have potential for use in the treatment of diabetesand obesity (Huang et al., Obes Rev. 10:610-616(2009)).

WO 2007/071402 describes the use of certain inner ionic phospholipids,phosphonolipids and phosphate derivatives for treatment or prevention ofallergic diseases. Furthermore, specific quaternary ammonium compoundsare disclosed in U.S. Pat No. 5,545,667 and U.S. Pat. No. 6,136,857 tobe useful as antineoplastic agents. Coy EA et al. Int Jimmunopharrnacol. 1990; 12(8):871-81 report a generalizedantiproliferative activity of specific amphiphilic molecules onT-Iymphocytes and on a variety of tumor cell lines and a lack ofspecificity for the immune system. WO 2009/136396 relates to certainsulfobetaines to be used in the treatment of cancer, obesity,age-related macular degeneration and neurodegenerative diseases. WO92/16201 relates to the use of specific betaine compounds for thetreatment of certain viral infections. Yan L, et al. Bioorg Med ChemLett 2004; 14(19):4861-6 describe certain aminopropyl-phosphonic acidderivatives as agonists of sphingosine-1-phosphate G protein-coupledreceptors, Birnie CR, et al. Antimicrob Agents Chemother. 2000;44(9):2514-2517 refer to certain N-alkyl-N,N-dimethyl betaines which arereported to show antimicrobial activity. Chen CKM, et al. J Med Chem.2008; 51(18):55945607 report the X-ray structures of certainbisphosphonates which comprise two phosphonate groups and are thusdifferent from the compounds of the present invention for at least thatreason. Rachinskii FY, et al. Journal of Applied Chemistry of USSR.1968; 41(10):2205-2207 describe specific N-carboxymethyl-azepanederivatives which do not have any substituents on the azepane ringcarbon atoms, and report bactericidal activity of these compounds. Thecompound having the CAS number 23035-15-6 (i.e.,1-(carboxymethyl)hexahydro-1-octadecyl-1H-azepinium chloride) does nothave any substituents on its azepane ring carbon atoms either. Thecompound having the CAS number 761356-67-6 (i.e.,3-methyl-4-(sulfomethyl)-1-(3-sulfopropyl)-1-tetradecylpyrrolidiniuminner salt) comprises a sulfomethyl substituent on its pyrrolidine ringand, for at least that reason, is different from the compounds of theinvention. Further specific quaternary ammonium compounds are, forexample, disclosed in: Ernst R et al. Toxicology. 1980; 15(3):233-42;Speijers GJ et al. Vaccine. 1989; 7(4):364-8; Vian L et al. Toxic inVitro. 1995; 9(2):185-190; Parris N, et al. Journal of the American OilChemists' Society. 1973; 50(12):509-512; U.S. Pat. No. 3,432,408;U.S.Pat. No. 4,085.134; and CN 101456810 A.

It has surprisingly been found that the compounds of the presentinvention, in particular the compounds of formula 1, 2, 3, 4, 5 or 6 asdescribed and defined herein below, have an advantageously lowcytotoxicity. The present invention thus solves the problem of providingtherapeutic agents having a favorable toxicity profile which areeffective, inter alia, in the treatment, prevention or amelioration ofinflammatory, autoimmune and/or allergic disorders. Accordingly, thepresent invention provides a compound of the following formula 1

or a pharmaceutical acceptable salt, solvate or prodrug thereof for usein the treatment, prevention or amelioration of an inflammatory,autoimmune and/or allergic disorder, wherein the inflammatory,autoimmune and/or allergic disorder is selected from: psoriasis, atopicdermatitis (atopic eczema), contact dermatitis, xerotic eczema,seborrheic dermatitis, neurodermitis, dyshidrosis, discoid eczema,venous eczema, dermatitis herpetiformis (Duhring's Disease),autoeczematization, dermatomyositis, hyper-IgE (Buckley) syndrome.Wiskott-Aldrich syndrome, anaphylaxis, food allergy, allergic reactionsto venomous stings, acute urticarias, chronic urticarias, physicalurticarias including aquagenic urticaria, cholinergic urticaria, coldurticaria (chronic cold urticaria), delayed pressure urticaria,dermatographic urticaria, heat urticaria, solar urticaria, vibrationurticaria, adrenergic urticaria, urticaria angioedema, inflammatorybowel disease, Crohn's disease, ulcerative colitis, collagenous colitis,lymphocytic colitis, diversion colitis (diverticulitis), Behcet'ssyndrome, indeterminate colitis, celiac disease, irritable bowelsyndrome, post-operative ileus, eosinophilic gastroenteropathy,gastritis, chronic allergic rhinitis, seasonal allergic rhinitis(hay-fever), allergic conjunctivitis, chemical conjunctivitis, neonatalconjunctivitis, Sjogren syndrome, open-angle glaucoma, dry eye disease(DED; including, e.g., aqueous tear-deficient dry eye (ADDE), Sjogrensyndrome dry eye (SSDE), non-SSDE, or evaporative dry eye (EDE)),diabetic macular edema (or diabetic retinopathy), chronic obstructivepulmonary disease (COPD), allergic asthma, allergic bronchopulmonaryaspergillosis, hypersensitivity pneumonitis, lung fibrosis, rheumatoidarthritis, juvenile rheumatoid arthritis, ankylosing spondylitis,systemic lupus erythematosus (SLE), scleroderma, reactive arthritis,polymyalgia rheumatica, Guillain-Barre syndrome, Hashimoto'sthyroiditis, Grave's disease, temporal arteritis, primary biliarycirrhosis, sclerosing cholangitis, autoimmune hepatitis, or alopeciaareata.

R¹ is a C₁₀₋₂₀ hydrocarbon group.

R² is a C₁₋₄ alkyl group, and R³ is —H, a CM alkyl group or R³ isabsent.

Alternatively, R² and R³ are mutually linked to form a pyrrolidine ring,a piperidine ring or an azepane ring together with the nitrogen atom Xto which they are attached, wherein said pyrrolidine ring, saidpiperidine ring or said azepane ring is optionally substituted with oneor more groups independently selected from —OH, —O(C₁₋₃ alkyl),—O—C(O)-(C₁₋₃alkyl), alkyl, —C(O)—(C₁₋₃ alkyl), —C(O)—NH₂,—C(O)—NH(C₁₋₃alkyl), —C(O)—N(C₁₋₃ alkyl)(C₁₋₃ alkyl), —NH₂, —NH(C₁₋₃alkyl), —N(C₁₋₃ alkyl)(C₁₋₃ alkyl), —NH—C(O)—(C₁₋₃ alkyl), —N(C₁₋₃alkyl)—C(O)—(C₁₋₃ alkyl), —NH—C(O)—O(C₁₋₃ alkyl) or —N(C₁₋₃alkyl)—C(O)—O(C₁₋₃ alkyl).

R⁴ is a C₁₋₆ alkylene group.

R⁵ is —SO₃, —SO₃H, —PO₃ ²⁻, —PO₃H₂, —PO₂(OC₁₋₃ alkyl),—PO₂H(OC₁₋₃alkyl), —PO(OC₁₋₃ alkyl)₂, —CO₂, —CO₂H or —CO₂(C₁₋₃ alkyl).

X is N⁺or, if R³ is absent, X is N.

The present invention further relates to a pharmaceutical compositioncomprising a compound of formula 1, as described and defined herein, ora pharmaceutical acceptable salt, solvate or prodrug thereof, and apharmaceutical acceptable exciplent for use in the treatment, preventionor amelioration of an inflammatory, autoimmune and/or allergic disorderselected from: psoriasis, atopic dermatitis (atopic eczema), contactdermatitis, xerotic eczema, seborrheic dermatitis, neurodermitis,dyshidrosis, discoid eczema, venous eczema, dermatitis herpetiformis(Duhring's Disease), autoeczematization, dermatomyositis, hyper-lgE(Buckley) syndrome. Wiskott-Aldrich syndrome, anaphylaxis, food allergy,allergic reactions to venomous stings, acute urticarias, chronicurticarias, physical urticarias including aquagenic urticaria,cholinergic urticaria, cold urticaria (chronic cold urticaria), delayedpressure urticaria, dermatographic urticaria, heat urticaria, solarurticaria, vibration urticaria, adrenergic urticaria, urticariaangioedema, inflammatory bowel disease, Crohn's disease, ulcerativecolitis, collagenous colitis, lymphocytic colitis, diversion colitis(diverticulitis). Behcet's syndrome, indeterminate colitis, celiacdisease, irritable bowel syndrome, post-operative ileus, eosinophilicgastroenteropathy, gastritis, chronic allergic rhinitis, seasonalallergic rhinitis (hay-fever), allergic conjunctivitis, chemicalconjunctivitis, neonatal conjunctivitis, Sjogren syndrome, open-angleglaucoma, dry eye disease (DED; including, e.g., aqueous tear-deficientdry eye (ADDE). Sjogren syndrome dry eye (SSDE), non-SSDE, orevaporative dry eye (EDE)), diabetic macular edema (or diabeticretinopathy), chronic obstructive pulmonary disease (COPD), allergicasthma, allergic bronchopulmonary aspergillosis, hypersensitivitypneumonitis, lung fibrosis, rheumatoid arthritis, juvenile rheumatoidarthritis, ankylosing spondylitis, systemic lupus erythematosus (SLE),scleroderma, reactive arthritis, polymyalgia rheum atica, Guillain-Barresyndrome, Hashimoto's thyroiditis, Grave's disease, temporal arteritis,primary biliary cirrhosis, sclerosing cholangitis, autoimmune hepatitis,or alopecia areata.

Moreover, the present invention relates to a method of treating,preventing or ameliorating an inflammatory, autoimmune and/or allergicdisorder, the method comprising the administration of a compound offormula 1, as described and defined herein, or a pharmaceuticallyacceptable salt, solvate or prodrug thereof, or a pharmaceuticalcomposition comprising any of the aforementioned entities and apharmaceutically acceptable excipient, to a subject (preferably, a humanor a non-human mammal) in need of such a treatment, prevention oramelioration, wherein the inflammatory, autoimmune and/or allergicdisorder is selected from: psoriasis, atopic dermatitis (atopic eczema),contact dermatitis, xerotic eczema, seborrheic dermatitis,neurodermitis, dyshidrosis, discoid eczema, venous eczema, dermatitisherpetiformis (Duhring's Disease), autoeczematization, dermatomyositis,hyper-IgE (Buckley) syndrome, Wiskott-Aldrich syndrome, anaphylaxis,food allergy, allergic reactions to venomous stings, acute urticarias,chronic urticarias, physical urticarias including aquagenic urticaria,cholinergic urticaria, cold urticaria (chronic cold urticaria), delayedpressure urticaria, dermatographic urticaria, heat urticaria, solarurticaria, vibration urticaria, adrenergic urticaria, urticariaangioedema, inflammatory bowel disease, Crohn's disease, ulcerativecolitis, collagenous colitis, lymphocytic colitis, diversion colitis(diverticulitis). Behcet's syndrome, indeterminate colitis, celiacdisease, irritable bowel syndrome, post-operative ileus, eosinophilicgastroenteropathy, gastritis, chronic allergic rhinitis, seasonalallergic rhinitis (hay-fever), allergic conjunctivitis, chemicalconjunctivitis, neonatal conjunctivitis, Sjögren syndrome, open-angleglaucoma, dry eye disease (DED; including, e.g., aqueous lear-deficienldry eye (ADDE), Sjogren syndrome dry eye (SSDE), non-SSDE, orevaporative dry eye (EDE)), diabetic macular edema (or diabeticretinopathy), chronic obstructive pulmonary disease (COPD), allergicasthma, allergic bronchopulmonary aspergillosis, hypersensitivitypneumonitis, lung fibrosis, rheumatoid arthritis, juvenile rheumatoidarthritis, ankylosing spondylitis, systemic lupus erythematosus (SLE),scleroderma, reactive arthritis, polymyalgia rheumatica, Guillain-Barresyndrome, Hashimoto's thyroiditis, Grave's disease, temporal arteritis,primary biliary cirrhosis, sclerosing cholangitis, autoimmune hepatitis,or alopecia areata.

Allergic and inflammatory responses are characterized by dynamicinteractions of immune and non-immune cells, coordinated throughcell-cell contact and soluble immune mediators. These responses andtheir outcomes are further modified by each individual's genetics andlifestyle.

T helper cells play a key role in initiation and maintenance ofinflammatory responses and can be divided into Th1 (cell-mediatedimmunity) and Th2 (antibody-mediated immunity) driven processes.Imbalances in these responses can result in pathological hyper- orhyposensitivity to antigens. A chronic inflammation manifests (n diversedisease states such as, for instance, inflammatory bowel disease,rheumatoid arthritis, atopic dermatitis, urticaria and psoriasis.

Inflammatory responses to antigens can take the form of helper T celldriven responses of different types. Th1 cells mediate cellularresponses involving cytotoxic cells such as macrophages, neutrophils andeosinophils, whereas Th2 cells mediate humoral responses involvingsecretion of antibodies from B cells and activation of mast cells. Othernon-immune responses, such as those involving cyclooxygenase andlipoxygenase may also be involved. Uncontrolled release of cytokines andchemokines is at the heart of inflammatory diseases, like inflammatorybowel disease, rheumatoid arthritis, atopic dermatitis, urticaria andpsoriasis.

A new interventional strategy is provided by the compounds according tothe present invention, in particular the compounds of formula 1, 2, 3,4, 5 or 6 as described and defined herein, which broadly modulate theactivities of proteins within the inflammatory cascade. Throughenrichment of the drug in membrane domains, an allosteric inhibition isexerted on key target proteins in signal transduction cascades ininflammation.

The compounds of the present invention were identified as potentinhibitors of immune mediator release in vitro in a mast cells model, asalso demonstrated in Example 27. Furthermore, they inhibited release ofthe proinflammatory cytokines, TNF-a and interleukin-6 from peripheralblood mononuclear cells (PBMCs) stimulated with lipopolysaccharide,demonstrating immunomodulatory activity in different cell types.

Broad anti-inflammatory activity was confirmed in animal models of Th1and Th2 driven inflammation. In a predominantly Th1-driven delayed typehypersensitivity (DTH) model in mice, the compounds suppressed theinflammatory response to an extent equivalent to dexamethasone, amarketed corticosteroid characterized by severe side effects, as shownin Example 29. In a predominantly Th2 driven allergic contact dermatitismodel, the compounds were highly active after topical application, asshown in Example 30, and also showed an anti-inflammatory effect afteroral administration.

In the context of the present invention, it was surprisingly found thatthe compounds of formula 1, 2, 3, 4, 5 or 6 as described and definedherein are potent inhibitors of mast cell degranulation and thusfunction as mast cell stabilizers and/or potent inhibitors of allergicand/or cellular inflammation. In particular, it was surprisingly foundthat the compounds as disclosed herein can be used therapeutically inthe treatment, prevention and/or amelioration of immunological disordersand disorders related to allergic and/or cellular inflammation, inparticular inflammatory, autoimmune and/or allergic disorders.

T helper (Th) cells are a subgroup of lymphocytes that play an importantrole in the immune system due to their participation in activating anddirecting other immune cells. The other major types of lymphocytes are Bcells and natural killer (NK) cells. During the antigenic activation andproliferation of Th cells, the Th0 cells differentiate into Th1, Th2 orother subtypes depending on the type of antigen, the antigen presentingcell and the cytokine environment.

Delayed type hypersensitivity, also called type IV hypersensitivity isan antibody-independent Th cell-mediated immune memory responseresulting from an over-stimulation of immune cells, commonly lymphocytesand macrophages, resulting in chronic inflammation and cytokine release.Important disease examples are contact dermatitis, chronic inflammationof ileum and colon, e.g. as seen in inflammatory bowel disease (IBD),rheumatoid arthritis and related diseases, ankylosing spondylitis,systemic lupus erythematosus, scleroderma, Gaucher's disease,fibromyalgia, osteoarthritis, reactive arthritis, pelvic inflammatorydisease, polymyalgia rheumatica, multiple sclerosis, Guillain-Barresyndrome, Hashimoto's thyroiditis, Grave's disease and chronictransplant rejection. For IBD, for instance. Hue et al. demonstrated acausal relationship between the disease and T cell-mediated intestinalinflammation (Hue, S; et al. (2006) J. Exp. Med. 203 (11), 2473).

Psoriasis is a chronic autoimmune disease affecting the skin. Onehypothesis for the cause of psoriasis sees the disease as being animmune-mediated disorder in which the excessive reproduction of skincells is secondary to factors produced by the immune system. T cellsbecome active, migrate to the dermis and trigger the release ofcytokines which cause inflammation and the rapid production of skincells.

Mast cells, or mastocytes, play a key role in the inflammatory process.When activated, the mast cell rapidly releases its characteristicgranules and various hormonal mediators into the the interstitium, aprocess called degranulation. The molecules released into theextracellular environment include preformed mediators, e.g. histamineand serotonin, newly formed lipid mediators (eicosanoids) and cytokines.In allergic reactions, mast cells remain inactive until an allergenbinds to the IgE receptor expressed at the cell surface, leading todegranulation and release of mediators.

Many forms of cutaneous and mucosal allergies, in most cases accompaniedby inflammatory symptoms, are mediated largely by mast cells. They playa central role in asthma, eczema, itch and the various forms ofrhinitis, conjunctivitis and urticaria. Mast cells are also implicatedin the pathology associated with disorders such as rheumatoid arthritis,bullous pemphigoid and multiple sclerosis. They have been shown to beinvolved in the recruitment of inflammatory cells to the joints andskin. Moreover, mastocytosis is a disorder featuring proliferation ofmast cells and exists in a cutaneous and systemic form.

Atopic dermatitis, also known as neurodermitis, is an inflammatory andpruritic skin disorder characterised by chronic inflammation. Althoughthe causes underlying atopic dermatitis are not well understood and therelationships between intake of, or contact with, allergens and variousinflammatory stimuli are not well established, it is postulated thatmast cell and/or T cell-related processes are involved in thepathological processes leading to atopic dermatitis.

Asthma and chronic obstructive pulmonary disease (COPD) are bothobstructive airway disorders, but differing types of inflammation areinvolved in the pathogenesis of these diseases. Asthma is frequently anallergic process with a preponderance of Th2 cells and eosinophils inthe airways. In contrast, there is predominant Th1 activity in the bloodof COPD patients (Lecki, M J; et al. (2003) Thorax 58, 23).

Dry eye disease (DED) is an inflammatory disorder of the lacrimalfunctional unit leading to chronic ocular surface disease, impairedquality of vision, and a wide range of complications. It is recognizedthat a chronic inflammatory response plays a key role in thepathogenesis of human dry eye disease (Calonge M, et al. Ocul ImmunolInflamm. 2010. 18:244-253; Stevenson W, et at. Arch Ophthalmol. 2012.130:90-100; Zoukhri D. Exp Eye Res. 2006. 82:885-898; Pflugfelder SC. AmJ Ophthalmol. 2004. 137:337-342).

Diabetic macular edema (or diabetic retinopathy) is characterized byearly retinal microvascular dysfunction and is a leading cause ofblindness in subjects suffering from diabetes. There is evidenceindicating that retinal inflammation plays an important role in thepathogenesis of diabetic macular edema (Joussen A M, et al. FASEBJ.2004. 18:1450-1452; Rangasamy S, et al. Middle East Afr J Ophthalmol.2012. 19:52-59; Meleth A D, et al. Invest Ophthalmol Vis Sci. 2005.46:4295-4301; Funatsu H, et al. Ophthalmology. 2009. 116:73-79; Kim S J,et al. Surv Ophthalmol. 2010. 55:108-133).

Accordingly, the compounds of the present invention, in particular thecompounds of formula 1, 2, 3, 4, 5 or 6, are useful in the treatment,prevention or amelioration of an inflammatory, autoimmune and/orallergic disorder.

The inflammatory, autoimmune and/or allergic disorder to be treated,prevented or ameliorated using the compounds of formula 1 or 2 accordingto the invention is selected from: psoriasis, atopic dermatitis (atopiceczema), contact dermatitis, xerotic eczema, seborrheic dermatitis,neurodermitis, dyshidrosts, discoid eczema, venous eczema, dermatitisherpetiformis (Duhring's Disease), autoeczematization, dermatomyositis,hyper-IgE (Buckley) syndrome, Wiskott-Aldrich syndrome, anaphylaxis,food allergy, or allergic reactions to venomous stings; acuteurticarias, chronic urticarias, physical urticarias including aquagenicurticaria, cholinergic urticaria, cold urticaria (chronic coldurticaria), delayed pressure urticaria, dermatographic urticaria, heaturticaria, solar urticaria, vibration urticaria, adrenergic urticaria,or urticaria angioedema; inflammatory bowel disease, Crohn's disease,ulcerative colitis, collagenous colitis, lymphocytic colitis, diversioncolitis (diverticulitis), Behcet's syndrome, indeterminate colitis,celiac disease, irritable bowel syndrome, post-operative ileus,eosinophilic gastroenteropathy, or gastritis; chronic allergic rhinitis,seasonal allergic rhinitis (hay-fever), allergic conjunctivitis,chemical conjunctivitis, neonatal conjunctivitis. Sjogren syndrome,open-angle glaucoma, dry eye disease (DED; including, e.g., aqueoustear-deficient dry eye (ADDE), Sjogren syndrome dry eye (SSDE),non-SSDE, or evaporative dry eye (EDE)), or diabetic macular edema (ordiabetic retinopathy); chronic obstructive pulmonary disease (COPO),allergic asthma, allergic bronchopulmonary aspergillosis,hypersensitivity pneumonitis, or lung fibrosis; rheumatoid arthritis,juvenile rheumatoid arthritis, ankylosing spondylitis, systemic lupuserythematosus (SLE), scleroderma, reactive arthritis, or polymyalgiarheumatica; or Guillain-Barre syndrome, Hashimoto's thyroiditis, Grave'sdisease, temporal arteritis, primary biliary cirrhosis, sclerosingcholangitis, autoimmune hepatitis, or alopecia areata.

The efficacy of the compounds of the present invention, in particularthe compounds of formula 1, 2, 3, 4, 5 or 6, in the inhibition of Aktkinase activation has furthermore been demonstrated in Example 28.

The compounds of the invention, which have been demonstrated to exhibitan efficacy in the suppression of the inflammatory response at leastequivalent to that of the corticosteroid dexamethasone, as also shown inExample 29, are furthermore advantageous in that they do not show theadverse effects which are usually observed for corticosteroids, such asreduction of lymph node weight and cell number (Example 30), which makesthem particularly useful in the treatment, prevention or amelioration ofinflammatory, autoimmune and/or allergic disorders.

Moreover, the compounds of the present invention, including thecompounds of formula 1, 2, 3, 4, 5 or 6, have a particularly lowcytotoxicity and, thus, an advantageous toxicity profile, as alsodemonstrated in Example 27.

The compound of formula 1 as defined above is described in more detailin the following.

R¹ is a C₁₀₋₂₀ hydrocarbon group. Preferably, R¹ is an alkyl group, analkenyl group, or an alkynyl group; more preferably, R¹ is a linearalkyl group, a linear alkenyl group, or a linear alkynyl group; evenmore preferably, R¹ is a linear alkyl group. The number of carbon atomsof the hydrocarbon group, the alkyl group, the alkenyl group, or thealkynyl group is 10 to 20, preferably 12, 14 or 16. Accordingly, it isparticularly preferred that R1is —(CH₂)₁₁—CH₃, —(CH₂)₁₃ —CH₃, or—(CH₂)₁₅—CH₃.

R² is a C₁₋₄ alkyl group, and R³ is —H, a alkyl group or R³ is absent;or R² and R³ are mutually linked to form a pyrrolidine ring, apiperidine ring or an azepane ring together with the nitrogen atom X towhich they are attached, wherein said pyrrolidine ring, said piperidinering or said azepane ring is optionally substituted with one or moregroups independently selected from —OH, —O(C₁₋₃ alkyl), —O—C(O)—(C₁₋₃alkyl), C₁₋₃ alkyl, —NH₂, —NH(C₁₋₃ alkyl) or —N(C₁₋₃ alkyl)(C₁₋₃ alkyl).

In one preferred embodiment, R² is methyl, and R³ is —H, a C₁₋₄ alkylgroup or R³ is absent. More preferably, R² is methyl, and R³ is —H,methyl or R³ is absent. Even more preferably, R² is methyl and R³ ismethyl.

In another preferred embodiment, R² and R³ are mutually linked to form apyrrolidine ring, a piperidine ring or an azepane ring together with thenitrogen atom X to which they are attached. More preferably, R² and R³are mutually linked to form a piperidine ring together with the nitrogenatom X to which they are attached. The pyrrolidine ring, the piperidinering or the azepane ring may be substituted with one or more (such as,e.g., one, two, three, or four), preferably one or two, more preferablyone, groups independently selected from —OH, —O(C₁₋₃ alkyl),—O—C(O)—(C₁₋₃ alkyl), C₁₋₃ alkyl, —C(O)—(C₁₋₃ alkyl), —C(O)—NH₂,—C(O)—NH(C₁₋₃ alkyl), —C(O)—N(C₁₋₃ alkyl)(C₁₋₃ alkyl), —NH₂, —NH(C₁₋₃alkyl), —N(C₁₋₃ alkyl)(C₁₋₃ alkyl), —NH—C(O)—(C₁₋₃ alkyl), —N(C₁₋₃alkyl)—C(O)—(C₁₋₃ alkyl), —NH-C(O)—O(C₁₋₃ alkyl) or —N(C₁₋₃ alkyl—C(O)—O9C₁₋₃ alkyl). Preferably, the pyrrolidine ring, the piperidinering or the azepane ring is unsubstituted or substituted with one ormore (such as, e.g., one, two, three, or four), preferably one or two,more preferably one, groups independently selected from —OH, —O(C₁₋₃alkyl), —O—C(O)—(C₁₋₃ alkyl), C₁₋₃ alkyl, —C(O)—NH₂, —C(O)—NH(C₁₋₃alkyl), —C(O)—N(C₁₋₃ alkyl)(C₁₋₃ alkyl), —NH₂, —NH(C₁₋₃ alkyl), or—N(C₁₋₃ alkyl)(C₁₋₃ alkyl), preferably selected independently from —OH,—O(C₁₋₃ alkyl), —O—C(O)—(C₁₋₃ alkyl), C₁₋₃ alkyl, —NH₂, —NH(C₁₋₃ alkyl),or —N(C₁₋₃ alkyl) (C₁₋₃ alkyl). More preferably, the pyrrolidine ring,the piperidine ring or the azepane ring is unsubstituted or substitutedwith one group selected from —OH, —O(C₁₋₃ alkyl), —O—C(O)—(C₁₋₃ alkyl),C₁₋₃ alkyl, —C(O)—NH₂, —C(O)—NH(C₁₋₃ alkyl), —C(O)—N(C₁₋₃ alkyl)(C₁₋₃alkyl), —NH₂, —N(C₁₋₃ alkyl) or —N(C₁₋₃ alkyl)(C₁₋₃ alkyl). Even morepreferably, the pyrrolidine ring, the piperidine ring or the azepanering is unsubstituted or substituted with one group —OH.

Accordingly, it is particularly preferred that R² and R³ are mutuallylinked to form a piperidine ring together with the nitrogen atom X towhich they are attached, wherein the piperidine ring is optionallysubstituted with one group —OH, preferably in para position with respectto the nitrogen atom X.

R⁴ is a C₁₋₆ alkylene group. The alkylene group may be linear orbranched; preferably, the aikylene group is linear. The alkylene grouphas 1 to 6 (i.e., 1, 2, 3, 4, 5 or 6) carbon atoms and preferably has 2or 3 carbon atoms, more preferably 3 carbon atoms. It is particularlypreferred that RA is —(CH₂)₃—.

R⁵ is —SO₃ ⁻, —SO₃H, —PO₃H⁻, —PO₃ ²⁻, —PO₃H₂, —PO₂(OC₁₋₃ alkyl)⁻,—PO₂H(OC₁₋₃ alkyl), —PO(OC₁₋₃ alkyl)₂, —CO₂ ⁻, —CO₂H or —CO₂(C₁₋₃alkyl). Preferably, R⁵ is −SO₃ ⁻, —SO₃H, −PO₃H⁻, —PO₃ ²⁻, or —PO₃H₂. Inone preferred embodiment, R⁵ is —SO₃ ⁻ or —SO₃H. In another preferredembodiment, R⁵ is —PO₃ ²⁻, —PO₃H or —PO₃H₂.

Accordingly, in a particularly preferred embodiment R⁵ is —SO₃ ⁻ or—SO₃H and R⁴ is a linear or branched alkylene group (preferably, R⁴ is alinear alkylene group having 2, 3, 4, 5 or 6 carbon atoms; morepreferably, R⁴ is a linear alkylene group having 3 carbon atoms; evenmore preferably, R⁴ is —(CH₂)³⁻). In a further particularly preferredembodiment R⁵ is —PO₃ ²⁻, —PO₃H or —PO₃H₂ and R⁴ is a linear or branchedC₂₋₆ alkylene group (preferably, R⁴ is a linear alkylene group having 2,3, 4, 5 or 6 carbon atoms; more preferably. R⁴ is a linear alkylenegroup having 3 carbon atoms; even more preferably, R⁴ is —(CH₂)₃—).

X is N⁺or, if R³ is absent, X is N.

A person skilled in the art understands that, if the compound of formula1 is provided in solution, the protonation of the acid group R⁵ dependson the pH of the solution. For example, if R⁵ is —PO₃H⁻ it may bepresent as —PO₃H₂ in a more acidic environment or as —PO₃ ²⁻ in a morealkaline environment.

Likewise, a skilled person understands that, if the compound of formula1 is provided in solution and if R³ in formula 1 is —H or is absent, theprotonation of the nitrogen atom X and, accordingly, the charge at thenitrogen atom X depends on the pH of the solution. Thus, depending onthe pH of the solution, R³ may be —H and X may be N⁺, or R³ may beabsent i and X may be N.

Preferred examples of the compound of formula 1 are the compounds 1a to1x shown below or pharmaceutical acceptable salts, solvates or prodrugsthereof:

In one embodiment described above, R² and R³ in formula 1 are mutuallylinked to form a pyrrolidine ring, a piperidine ring or an azepane ringtogether with the nitrogen atom X to which they are attached, whereinsaid pyrrolidine ring, said piperidine ring or said azepane ring isoptionally substituted with one or more groups independently selectedfrom —OH, —O(C₁₋₃ alkyl), —O—C(O)—(C₁₋₃alkyl), C₁₋₃ alkyl, —C(O)—(C₁₋₃alkyl), —C(O)—NH₂, —C(O)—NH(C₁₋₃alkyl), —C(O)—N(C₁₋₃ alkyl)(C₁₋₃ alkyl),—NH₂, —NH(C₁₋₃ alkyl), —N(C₁₋₃ alkyl)(C₁₋₃ alkyl), —NH—C(O)—(C₁₋₃alkyl), —N(C₁₋₃ alkyl)—C(O)—(C₁₋₃ alkyl), —NH—C(O)—O(C₁₋₃ alkyl) or—N(C₁₋₃ alkyl) —C(O)—O(C₁₋₃ alkyl).

Accordingly, the compound of formula 1 may be a compound of thefollowing formula 2

or a pharmaceutical acceptable salt, solvate or prodrug thereof for usein the treatment, prevention or amelioration of an inflammatory,autoimmune and/or allergic disorder.

In formula 2, the groups R¹, R⁴, and R⁵ have the meanings or thepreferred meanings defined herein above for the compound of formula 1.

n is 1, 2, or 3. Preferably, n is 2.

m is an integer from 0 to 4. Preferably, m is 0, 1, or 2; morepreferably, m is 0 or 1; even more preferably, m is 1.

Each R⁶ is independently selected from —OH, —O(C,.3 alkyl),—O—C(O)—(C₁₋₃ alkyl), C₁₋₃ alkyl, —C(O)—(C₁₋₃ alkyl), —C(O)—NH₂,—C(O)—NH(C₁₋₃ alkyl), —C(O)—N(C₁₋₃ alkyl)(C₁₋₃ alkyl), —NH₂, —NH(Ci-3alkyl), —N(C₁₋₃ alkyl)(C₁₋₃ alkyl), —NH—C(O)—(C₁₋₃ alkyl), —N(C₁₋₃alkyl)—C(O)—(C₁₋₃ alkyl). —NH—C(O)—O(C₁₋₃ alkyl) or —N(C₁₋₃alkyl)—C(O)—O(C₁₋₃ alkyl). Preferably, each R⁶ is independently selectedfrom —OH, —O(C₁₋₃ alkyl), —O—C(O)—(C₁₋₃ alkyl), C₁₋₃ alkyl, —C(O)—NH₂,—C(O)—NH(C₁₋₃ alkyl), —C(O)—N(C₁₋₃ alkyl)(C₁₋₃ alkyl), —NH₂, —NH(C₁₋₃alkyl), or alkyl)(C₁₋₃ alkyl). More preferably, each R⁶ is independentlyselected from —OH, —O(C₁₋₃ alkyl), —O—C(O)—(C₁₋₃ alkyl), C₁₋₃ alkyl,—NH₂, —NH(C₁₋₃ alkyl) or —N(C₁₋₃ alkyl)(C₁₋₃ alkyl). Even morepreferably, each R⁶ is —OH.

It is to be understood that each R⁶ is attached to a carbon atom of thepyrrolidine, piperidine or azepane ring. It is further to be understoodthat, if m is 0, the pyrrolidine, piperidine or azepane ring (to whichR⁶ would be attached) is unsubstituted, i.e. is substituted withhydrogen.

In one embodiment, n is 1 or 3, and m is 0,

In a preferred embodiment, n is 2, m is 1, and R⁶ is —OH, —O(C₁₋₃alkyl), —O—C(O)—(C₁₋₃ alkyl), C₁₋₃ alkyl, —NH₂, —NH(C₁₋₃ alkyl) or—N(C₁₋₃ alkyl)(C₁₋₃ alkyl), in particular —OH. In this embodiment, it isfurther preferred that R⁶ is in para position in respect of the ringnitrogen atom N⁺.

Preferred examples of the compound of formula 2 are the compounds 2a to2d shown below or pharmaceutically acceptable salts, solvates orprodrugs thereof:

The invention furthermore relates to a compound of formula 3 or apharmaceutical acceptable salt, solvate or prodrug thereof for use as amedicament.

In formula 3, the groups R¹ and X have the same meanings and preferredmeanings as described and defined herein above for the correspondinggroups in formula 1.

R² in formula 3 is a C₁₋₃ alkyl group, and R³ is —H or R² is absent; orR² and R³ are mutually linked to form a pyrrolidine ring, a piperidinering or an azepane ring together with the nitrogen atom X to which theyare attached, wherein said pyrrolidine ring, said piperidine ring orsaid azepane ring is substituted with one or more groups independentlyselected from —OH, —O(C₁₋₃ alkyl), —O—C(O₁₋₃ alkyl), alkyl, —C(O)—(C₁₋₃alkyl), —C(O)—NH₂, —C(O)—NH(C₁₋₃ alkyl), —C(O)—N(C₁₋₃ alkyl)(C₁₋₃alkyl), —NH₂, —NH(C₁₋₃ alkyl), —N(C₁₋₃ alkyl)(C₁₋₃ alkyl),—NH—C(O)—(C₁₋₃ alkyl), —N(C₁₋₃ alkyl)—C(O)—(C₁₋₃ alkyl), —NH—C(O)—O(C₁₋₃alkyl) or —N(C₁₋₃ alkyl)—C(O)—O(C₁₋₃ alkyl).

In one preferred embodiment, R² is methyl, and R³ is —H or R³ is absent.

In another preferred embodiment, R² and R³ are mutually linked to form apyrrolidine ring, a piperidine ring or an azepane ring together with thenitrogen atom X to which they are attached. More preferably, R² and R³are mutually linked to form a piperidine ring together with the nitrogenatom X to which they are attached. The pyrrolidine ring, the piperidinering or the azepane ring is substituted with one or more (such as, e.g.,one, two, three, or four), preferably one or two, more preferably one,groups independently selected from —OH, —O(C₁₋₃ alkyl), —O—C(O)—(C₁₋₃alkyl), C₁₋₃ alkyl, —C(O)—(C₁₋₃ alkyl), —C(O)—NH₂, —C(O)—NH(C₁₋₃ alkyl),—C(O)—N(C₁₋₃ alkyl)(C₁₋₃ alkyl), —NH₂, —NH(C₁₋₃ alkyl), —N(C₁₋₃alkyl)(C₁₋₃ alkyl), —NH—C(O)—(C₁₋₃ alkyl), —N(C₁₋₃ alkyl)—C(O)—(C₁₋₃alkyl), —NH—C(O)—O(C₁₋₃ alkyl) or —N(C₁₋₃ alkyl)—C(O)—O(C₁₋₃ alkyl).

Preferably, the pyrrolidine ring, the piperidine ring or the azepanering is substituted with one or more (such as, e.g., one, two, three, orfour), preferably one or two, more preferably one, groups independentlyselected from —OH, —O(C₁₋₃ alkyl), —O—C(O)—(C₁₋₃ alkyl), C₁₋₃ alkyl,—C(O)—NH₂, —C(O)—NH(C₁₋₃ alkyl), —C(O)—N(C₁₋₃ alkyl)(C₁₋₃ alkyl),—NH₂,—NH(C₁₋₃ alkyl), or —N(C₁₋₃ alkyl)(C₁₋₃ alkyl), preferably selectedindependently from —OH, —O(C₁₋₃ alkyl), —O—C(O)—(C₁₋₃ alkyl), C₁₋₃alkyl, —NH₂,—NH(C₁₋₃ alkyl), or —N(C₁₋₃ alkyl)(C₁₋₃ alkyl). Morepreferably, the pyrrolidine ring, the piperidine ring or the azepanering is substituted with one group selected from —OH, —O(C₁₋₃ alkyl),—O—C(O)—(C₁₋₃ alkyl), C₁₋₃ alkyl, —C(O)—NH₂, —C(O)—NH(C₁₋₃ alkyl),—C(O)—N(C₁₋₃ alkyl)(C₁₋₃ alkyl), —NH₂, —NH(C₁₋₃ alkyl), or —N(C₁₋₃alkyl)(C₁₋₃ alkyl). Even more preferably, the pyrrolidine ring, thepiperidine ring or the azepane ring is substituted with one group —OH.Accordingly, it is particularly preferred that R² and R³ are mutuallylinked to form a piperidine ring together with the nitrogen atom X towhich they are attached, wherein the piperidine ring is substituted withone group —OH, preferably in para position with respect to the nitrogenatom X.

R⁴ is a C₁₋₆ alkylene group. The alkylene group may be linear orbranched; preferably, the alkylene group is linear. The alkylene grouphas 1 to 6 (i.e., 1, 2, 3, 4, 5 or 6) carbon atoms and preferably has 2or 3 carbon atoms, more preferably 3 carbon atoms. It is particularlypreferred that R⁴ is —(CH₂)₃—.

If R⁴ is an alkylene group having 1, 2, 4, 5 or 6 carbon atoms, then R⁵is —SO₃, —SO₃H, —PO₃H⁻, —PO₃ ²⁻, —PO₃H₂, —PO₂(OC₁₋₃ alkyl)⁻, —PO₂H(OC₁₋₃alkyl), —PO(OC₁₋₃ alkyl)₂, —CO₂ ⁻, —CO₂H, or −CO₂(C₁₋₃ alkyl), andpreferably is R⁵ is —SO₃ ⁻, —SO₃H, —PO₃H⁻, —PO₃ ²⁻, or —PO₃H₂. If R⁴ isan alkylene group having 3 carbon atoms, then R⁵ is —SO₃, —SO₃H, —CO₂ ⁻,—CO₂H, —CO₂(C₁₋₃ alkyl), —PO₂(OC₁₋₃ alkyl), —PO(OC₁₋₃ alkyl)₂, or—PO(OC₁₋₃ alkyl)₂, and preferably R⁵ is —SO₃ ⁻, —SO₃H, —CO₂ ⁻or −CO₂H,more preferably —SO₃ ⁻ or —SO₃H.

Accordingly, in a particularly preferred embodiment R⁵ is —SO₃ or —SO₃Hand R⁴ is a linear or branched C₂₋₆ alkylene group (preferably, R⁴ is alinear alkylene group having 2, 3, 4, 5 or 6 carbon atoms; morepreferably, R⁴ is a linear alkylene group having 3 carbon atoms; evenmore preferably, R⁴ is —(CH₂)₃—). In a further particularly preferredembodiment R⁵ is —PO₃ ²⁻, —PO₃H⁻ or —PO₃H₂ and R⁴ is a linear orbranched alkylene group having 2, 4, 5 or 6 carbon atoms (preferably, R⁴is a linear alkylene group having 2, 4, 5 or 6 carbon atoms). In afurther particularly preferred embodiment, R⁴ is an alkylene grouphaving 1, 2, 4, 5 or 6 carbon atoms, and R⁵ is —PO₃H⁻, —PO₃ ²⁻, —PO₃H₂,—PO₂(OC₁₋₃ alkyl)⁻, —PO₂H(OC₁₋₃ alkyl), —PO(OC₁₋₃ alkyl)₂, —CO₂ ⁻,—CO₂H, or —CO₂(C₁₋₃ alkyl); or R⁴ is an alkylene group having 3 carbonatoms, and R⁵ is —CO₂ ⁻, —CO₂H, —CO₂(C₁₋₃ alkyl), —PO₂(OC₁₋₃ alkyl)⁻,—PO₂H(OC₁₋₃ alkyl), or —PO(OC₁₋₃ alkyl)₂ .

Preferred examples of the compound of formula 3 are the compounds 1b,1n, 1q, 1s, 1w, 1x, 2b, 2c or 2d shown below or pharmaceuticalacceptable salts, solvates or prodrugs thereof:

In accordance with the above definition of the compound of formula 3,the groups R² and R³ may, in one embodiment, be mutually linked to forma pyrrolidine ring, a piperidine ring or an azepane ring together withthe nitrogen atom X to which they are attached, wherein said pyrrolidinering, said piperidine ring or said azepane ring is substituted with oneor more groups independently selected from —OH, —O(O₁₋₃ alkyl),—O—C(O)—(C₁₋₃ alkyl), C₁₋₃ alkyl, —C(O)—(C₁₋₃ alkyl), —C(O)—NH₂,—C(O)—NH(C₁₋₃ alkyl), —C(O)—N(C₁₋₃ alkyl)(C₁₋₃ alkyl), —NH₂, —NH(C₁₋₃alkyl), —N(C₁₋₃ alkyl)(C₁₋₃ alkyl), —NH—C(O)—(C₁₋₃ alkyl), —N(C₁₋₃alkyl)—C(O)—(C₁₋₃ alkyl), —NH—C(O)—O(C₁₋₃ alkyl) or —N(C₁₋₃alkyl)—C(O)—O(C₁₋₃ alkyl).

The compound of formula 3, which is provided herein as a medicament, maythus be a compound of the following formula 4

or a pharmaceutically acceptable salt, solvate or prodrug thereof.

The groups R¹, R⁴ and R⁵ in formula 4 have the same meanings andpreferred meanings as defined herein above for ihe corresponding groupsin formula 3.

n is 1, 2, or 3. Preferably, n is 2.

In formula 4, m is an integer from 1 to 4. Preferably, m is 1 or 2; morepreferably, m is 1.

Each R⁶ is independently selected from —OH, —O(C₁₋₃ alkyl),—O—C(O)—(C₁₋₃ alkyl), C₁₋₃ alkyl, —C(O)—(C₁₋₃ alkyl), —C(O)—NH₂,—C(O)—NH(C₁₋₃ alkyl), —C(O)—N(C₁₋₃ alkyl)(C₁₋₃ alkyl), —NH₂, —NH(C₁₋₃alkyl), —N(C₁₋₃ alkyl)(C₁₋₃ alkyl). —NH—C(O)—(C₁₋₃ alkyl), —N(C₁₋₃alkyl)—C(O)—(C₁₋₃ alkyl), —NH—C(O)—O(C₁₋₃ alkyl) or —N(C₁₋₃alkyl)—C(O)—O(C₁₋₃ alkyl). Preferably, each R⁶ is independently selectedfrom —OH, —O(C₁₋₃ alkyl), —O—C(O)—(C₁₋₃ alkyl), C₁₋₃ alkyl, —C(O)—NH₂,—C(O)—NH(C₁₋₃ alkyl), —C(O)—N(C₁₋₃ alkyl)(C₁₋₃ alkyl), —NH₂, —NH(C₁₋₃alkyl), or —N(C₁₋₃ alkyl)(C₁₋₃ alkyl). More preferably, each R⁶ isindependently selected from —OH, —O(C₁₋₃ alkyl), —O—C(O)—(C₁₋₃ alkyl),C₁₋₃ alkyl, —NH₂, —NH(C₁₋₃ alkyl) or —N(C₁₋₃ alkyl)(C₁₋₃ alkyl). Evenmore preferably, each R⁶ is —OH.

It is to be understood that each R⁶ is attached to a carbon atom of thepyrrolidine, piperidine or azepane ring.

In a preferred embodiment, n in formula 4 is 2, m is 1, and R⁶ is —OH,—O(C₁₋₃ alkyl), —O—C(O)—(C₁₋₃ alkyl), C₁₋₃ alkyl, —NH₂, —NH(C₁₋₃ alkyl)or —N(C₁₋₃ alkyl)(C₁₋₃ alkyl), in particular —OH. In this embodiment, itis further preferred that R⁶ is in para position in respect of the ringnitrogen atom N⁺.

Preferred examples of the compound of formula 4 are the compounds 2b, 2cor 2d shown below or pharmaceutically acceptable salts, solvates orprodrugs thereof:

The present invention also relates to a pharmaceutical compositioncomprising a compound of formula 3 or 4, as described and definedherein, or a pharmaceutically acceptable salt, solvate or prodrugthereof, and a pharmaceutically acceptable excipient.

The invention further relates to a compound of formula 3 or 4, asdescribed and defined herein, or a pharmaceutically acceptable salt,solvate or prodrug thereof, or a pharmaceutical composition comprisingany of the aforementioned entities and a pharmaceutically acceptableexcipient, for use in the treatment, prevention or amelioration of aninflammatory, autoimmune and/or allergic disorder.

Moreover, the present invention relates to a method of treating,preventing or ameliorating a disease or disorder, in particular aninflammatory, autoimmune and/or allergic disorder, the method comprisingthe administration of a compound of formula 3 or 4, as described anddefined herein, or a pharmaceutically acceptable salt, solvate orprodrug thereof, or a pharmaceutical composition comprising any of theaforementioned entities and a pharmaceutically acceptable excipient, toa subject preferably, a human or a non-human mammal) in need of such atreatment, prevention or amelioration.

The inflammatory, autoimmune and/or allergic disorder to be treated,prevented or ameliorated using the compounds of formula 3 or 4 accordingto the invention is, for example, selected from: psoriasis, atopicdermatitis (atopic eczema), contact dermatitis, xerotic eczema,seborrheic dermatitis, neurodermitis, dyshidrosis, discoid eczema,venous -eczema, dermatitis herpetiformis (Duhring's Disease),autoeczematization, dermatomyasitis, hyper-IgE (Buckley) syndrome,Wiskott-Aldrich syndrome, anaphylaxis, food allergy, or allergicreactions to venomous slings; acute urticarias, chronic urticarias,physical urticarias including aquagenic urticaria, cholinergicurticaria, cold urticaria (chronic cold urticaria), delayed pressureurticaria, dermatographic urticaria, heat urticaria, solar urticaria,vibration urticaria, adrenergic urticaria, or urticaria angioedema;inflammatory bowel disease, Crohn's disease, ulcerative colitis,collagenous colitis, lymphocytic colitis, diversion colitis(diverticulitis), Behcet's syndrome, indeterminate colitis, celiacdisease, irritable bowel syndrome, post-operative ileus, eosinophilicgastroenteropathy, or gastritis; chronic allergic rhinitis, seasonalallergic rhinitis (hay-fever), allergic conjunctivitis, chemicalconjunctivitis, neonatal conjunctivitis, Sjögren syndrome, open-angleglaucoma, dry eye disease (DED; including, e.g., aqueous tear-deficientdry eye (ADDE). Sjögren syndrome dry eye (SSDE), non-SSDE, orevaporative dry eye (EDE)), or diabetic macular edema (or diabeticretinopathy); chronic obstructive pulmonary disease (COPD), allergicasthma, allergic bronchopulmonary aspergillosis, hypersensitivitypneumonitis, or lung fibrosis; rheumatoid arthritis, juvenile rheumatoidarthritis, ankylosing spondylitis, systemic lupus erythematosus (SLE),scleroderma, osteoarthritis, reactive arthritis, or polymyalgiarheumatica; multiple sclerosis, Guillain-Barre syndrome, Hashimoto'sthyroiditis, Grave's disease, temporal arteritis, primary biliarycirrhosis, sclerosing cholangitis, autoimmune hepatitis, alopecia areataor autoimmune lymphoproliferative syndrome (ALPS); a graft-versus-hostdisease, a host-versus-graft disease or a transplant rejection; or aninflammatory contribution to Alzheimer's disease or Parkinson's disease.

The present invention further provides novel compounds. These compoundsare described herein and are characterized by formula 5 or 6 as definedbelow. The compounds of formula 5 or 6 as provided in the context of thepresent invention are particularly useful In a medical setting, i.e. aspharmaceuticals. As is evident form the disclosure of the invention,these compounds are particularly useful in the treatment, prevention oramelioration of an inflammatory, autoimmune and/or allergic disorder.The invention thus further provides a method of treating, preventing orameliorating a disease or disorder, in particular an inflammatory,autoimmune and/or allergic disorder, the method comprising theadministration of a compound of formula 5 or 6 as defined below or apharmaceutical acceptable salt, solvate or prodrug thereof, or apharmaceutical composition comprising any of the aforementioned entitiesand a pharmaceutical acceptable excipient, to a subject (preferably, ahuman or a non-human mammal) in need of such a treatment, prevention oramelioration.

Accordingly, the invention provides a compound of the following formula5

or a pharmaceutical acceptable salt, solvate or prodrug thereof, whereinR¹, R² , R³ , and X have the meanings or the preferred meanings definedherein above for the compound of formula 3.

R⁴ in formula 5 is a C₁₋₆ alkylene group. The alkylene group may belinear or branched; preferably, the alkylene group is linear. Thealkylene group has 1 to 6 (i.e., 1, 2, 3, 4, 5 or 6) carbon atoms andpreferably has 2 or 3 carbon atoms, more preferably 3 carbon atoms. Itis particularly preferred that R⁴ is —(CH₂)₃ ⁻.

If R⁴ is an alkylene group having 1, 2, 4, 5 or 6 carbon atoms, then R⁵is —SO₃ ^(−, —SO) ₃H, —PO₃H —PO₃ ²⁻, —PO₃H₂, —PO₂(OC₁₋₃ alkyl)⁻,—PO₂H(OC₁₋₃ alkyl), —PO(OC₁₋₃ alkyl)⁻, —CO₂, —CO₂H, or —CO₂(C₁₋₃ alkyl),and preferably is R⁵ is —SO₃ ⁻, —SO₃H, —PO₃H —PO₃ ²⁻, or -P03H2. If R4is an alkylene group having 3 carbon atoms, then R⁵ is —CO₂, —CO₂H,—CO₂(C₁₋₃ alkyl), —PO₂(OC₁₋₃ alkyl)⁻, —PO₂H(OC₁₋₃ alkyl), or —PC(OC₁₋₃alkyl)₂, and preferably R₅ is —CO₂ ⁻ or —CO₂H. Accordingly, in aparticularly preferred embodiment R⁵ is —SO₃ or —SO₃H and R⁴ is a linearor branched alkylene group having 2, 4, 5 or 6 carbon atoms (preferably,R⁴ is a linear alkylene group having 2, 4, 5 or 6 carbon atoms). In afurther particularly preferred embodiment R⁵ is —PO₃ 2′, —PO₃H⁻, or—PO_(3 Hz and R) ⁴ is a linear or branched alkylene group having 2, 4, 5or 6carbon atoms (preferably, R⁴ is a linear alkylene group having 2, 4,5 or 6 carbon atoms). In a further particularly preferred embodiment, R4is an alkylene group having 1, 2, 4, 5 or 6 carbon atoms, and R⁵ is—PO₃H⁻, —PO₃ 2 ⁻, −PO₃H₂, -PO₂(OC₁₋₃ alkyl)⁻, −PO₂H(Oc₁₋₃ alkyl),—PO₂(OC₁₋₃ alkyl)2, —CO₂ , CO₂H, or —CO₂(C₁₋₃ alkyl); or R⁴ is analkylene group having 3 carbon atoms, and R⁵ is —CO₂ ⁻, —CO₂H, —CO₂(C₁₋₃alkyl), —PO₂(OC₁₋₃ alkyl)³¹, —PO₂H(OC₁₋₃ alkyl), or —PO(OC₁₋₃ alkyl)₂.Preferred examples of the compound of formula 5 are the compounds 1 n, 1q, 1 s, 1 w or 1 x shown below or pharmaceutically acceptable salts,solvates or prodrugs thereof:

Furthermore, the compound of formula 5 may be a compound of thefollowing formula 6

or a pharmaceutical acceptable salt, solvate or prodrug thereof, whereinR¹ , R⁴ and R⁵ have the meanings or the preferred meanings definedherein above for the compound of formula 5, and R⁶, n and m have themeanings or the preferred meanings defined herein above for the compoundof formula 4.

The compounds to be used in accordance with the present invention, inparticular the compounds of formula 1, 2, 3, 4, 5 or 6, can be preparedby methods known in the field of synthetic chemistry.

For example, compounds of the general formula 1 can be prepared bynucleophilic ring opening of alkanesultones with N-alkylatedalkylamines. Alternatively, compounds of the general formula 1 can beprepared by N-alkylation of N-alkylated alkylamines (or otherN-alkylated hydrocarbylamines, such as, e.g., N-alkylated alkenylaminesor N-alkylated alkynylamines) using ω-chloro- orω-bromo-1-alkanesulfonates under basic conditions. In a similar way, therelated phosphonates or carboxylates are generated, e.g., by usingω-chloro- or ω-bromo-1-alkanephosphonates or ω-chloro- or(ω-bromo-1-alkanecarboxyfates. The corresponding N,N-dialkylatedquarternary ammonium derivatives can be obtained by standardN-alkylation using, e.g., alkyl iodides. Compounds of formula 3 or 5 canbe prepared as described herein for the compounds of formula 1.Compounds of the general formula 2 (and, likewise, compounds of formula4 or 6) can be prepared by consecutive N-alkylation of appropriatelyfunctionalised pyrrolidine, piperidine or azepane derivatives using theaforementioned ω-chloro-1-alkanesulfonates, -phosphonates or-carboxylates followed by alkyl iodides.

The compounds of formula 1, 2, 3, 4, 5 or 6 can also be prepared Inanalogy to the synthetic routes described in the examples section.

As used herein, the term “hydrocarbon group” refers to a groupconsisting of carbon atoms and hydrogen atoms, which group may besaturated or unsaturated, linear, branched or cyclic, aliphatic oraromatic. A “C₁₀₋₂₀ hydrocarbon group” denotes a hydrocarbon grouphaving 10 to 20 carbon atoms.

As used herein, the term “alkyl group” refers to a monovalent saturatedaliphatic (i.e. non-aromatic) acyclic hydrocarbon group which may belinear or branched and does not comprise any carbon-to-carbon doublebond or any carbon-to-carbon triple bond.

As used herein, the term “alkenyl group” refers to a monovalentunsaturated aliphatic acyclic hydrocarbon group which may be linear orbranched and comprises at least one carbon-to-carbon double bond whileit does not comprise any carbon-to-carbon triple bond.

As used herein, the term “alkynyt group” refers to a monovalentunsaturated aliphatic acyclic hydrocarbon group which may be linear orbranched and comprises at least one carbon-to-carbon triple bond andoptionally one or more carbon-to-carbon double bonds.

As used herein, the term “alkylene group” refers to a divalent saturatedaliphatic (i.e. non-aromatic) acyclic hydrocarbon group which may belinear or branched and does not comprise any carbon-to-carbon doublebond or any carbon-to-carbon triple bond.

The scope of the invention embraces all pharmaceutical acceptable saltforms of the compounds of formula 1, 2, 3, 4, 5 or 6, which may beformed, e.g., by protonation of an atom carrying an electron lone pairwhich is susceptible to protonation, such as an amino group, with aninorganic or organic acid, or as a salt of a carboxylic acid group witha physiologically acceptable cation as they are well-known in the art.Exemplary base addition salts comprise, for example, alkali metal saltssuch as sodium or potassium salts; alkaline earth metal salts, such ascalcium or magnesium salts; ammonium salts; aliphatic amine salts, suchas trimethylamine, triethylamine, dicyclohexylamine, ethanolamine,diethanolamine, triethanolamine, procaine salts, meglumine salts,diethanol amine salts or ethylenediamine salts; aralkyl amine salts suchas N,N-dibenzylethylenediamine salts, benetamine salts; heterocyclicaromatic amine salts, such as pyridine salts, picofine salts, quinolinesalts or isoquinoline salts; quaternary ammonium salts, such astetramethylammonium salts, tetraethylammonium salts,benzyltrimethylamrnonium salts, benzyltriethylammonium salts,benzyltributylammonium salts, methyltrioctylammonium salts ortetrabutylammonium salts; and basic amino acid salts, such as argininesalts or lysine salts. Exemplary acid addition salts comprise, forexample, mineral acid salts, such as hydrochloride, hydrobromide,hydroiodide, sulfate salts, nitrate salts, phosphate salts (such asphosphate, hydrogenphosphate or dihydrogenphosphate salts), carbonatesalts, hydrogen carbonate salts or perchlorate salts; organic acid saltssuch as acetate, propionate, butyrate, pentanoate, hexanoate,heptanoate, octanoate, cyclop entaneprop ion ate, undecanoate, lactate,maleate, oxalate, fumarate, tartrate, malate, citrate, nicotinate,benzoate, salicylate or ascorbate salts; sulfonate salts, such asmethanesulfonate, ethanesuifonate, 2-hydroxyethanesulfonate,benzenesulfonate, p-toluenesulfonate (tosylate), 2-naphthaIenesulfonate,3-phenylsulfonate, or camphorsulfonate salts; and acidic amino acidsalts, such as aspartate or glutamate salts.

Moreover, the scope of the invention embraces solid forms of thecompounds of formula 1, 2, 3, 4, 5 or 6 in any solvated form, includinge.g. solvates with water, for example hydrates, or with organic solventssuch as, e.g., methanol, ethanol, isopropanol or acetonitrile, i.e. as amethanolate, ethanolate, isopropanolate or acetonitrilate, respectively;or in the form of any polymorph.

Furthermore, the formulae in the present application are intended tocover all possible stereoisomers, including enantiomers anddiastereomers, of the indicated compounds.

Thus, all stereoisomers of the compounds of the present invention, inparticular the compounds of formula 1, 2, 3, 4, 5 or 6, are contemplatedas part of the present invention, either in admixture or in pure orsubstantially pure form. The scope of the compounds according to theinvention embraces all the possible stereoisomers and their mixtures. Itvery particularly embraces the racemic forms and the isolated opticalisomers. The racemic forms can be resolved by physical methods, such as,e.g., fractional crystallization, separation or crystallization ofdiastereomeric derivatives or separation by chlral columnchromatography. The individual optical isomers can be obtained from theracemates using conventional methods, such as, e.g., salt formation withan optically active acid followed by crystallization.

Pharmaceutically acceptable prodrugs of compounds of the presentinvention, in particular of the compounds of formula 1, 2, 3, 4, 5 or 6,are derivatives which have chemically or metabolically cleavable groupsand become, by solvolysls or under physiological conditions, thecompounds of the present invention which are pharmaceutical active invivo. Prodrugs of compounds of the present invention may be formed in aconventional manner with a functional group of the compounds such aswith an amino, hydroxy or carboxy group. The prodrug derivative formoften offers advantages of solubility, tissue compatibility or delayedrelease in a mammalian organism (see, Bundgaard, H., Design of Prodrugs,pp. 7-9, 21-24, Elsevier, Amsterdam 1985). Prodrugs include acidderivatives well known to the person skilled in the art, such as, forexample, esters prepared by reaction of the parent acidic compound witha suitable alcohol, or amides prepared by reaction of the parent acidcompound with a suitable amine. When a compound of the present inventionhas a carboxyl group, an ester derivative prepared by reacting thecarboxyl group with a suitable alcohol or an amide derivative preparedby reacting the carboxyl group with a suitable amine is exemplified as aprodrug. An especially preferred ester derivative as a prodrug ismethylester, ethylester, n-propylester, isopropylester, n-butylester,isobutylester, tert-butylester, morpholinoethylester,N,N-diethylglycolamidoester or a-acetoxyethylester. When a compound ofthe present invention has a hydroxy group, an acyloxy derivativeprepared by reacting the hydroxyl group with a suitable acylhalide or asuitable acid anhydride is exemplified as a prodrug. An especiallypreferred acyloxy derivative as a prodrug is —OC(═O)—CH₃, —OC(═O)—C₂H₅,—OC(═O)—(tert-Bu), —OC(═O)‥C₁₅H₃₁, —OC(═O)—(m—COONa—Ph),—OC(═O)—CH₂CH₂COONa, —O(OO)—CH(NH₂)CH₃ or —OC(═O)—CH₂—N(CH₃)₂. When acompound of the present invention has an amino group, an amidederivative prepared by reacting the amino group with a suitable acidhalide or a suitable mixed anhydride is exemplified as a prodrug. Anespecially preferred amide derivative as a prodrug is —NHC(═O)—(CH₂)OCH₃or —NHC(═O)—CH(NH₂)CH₃.

The compounds described herein may be administered as compounds per sein their use as pharmacophores or pharmaceutical compositions or may beformulated as medicaments. Within the scope of the present invention arepharmaceutical compositions comprising as an active ingredient acompound of formula 1, 2, 3, 4, 5 or 6 as defined above. Thepharmaceutical compositions may optionally comprise one or morepharmaceutical acceptable excipients, such as carriers, diluents,fillers, disintegrants, lubricating agents, binders, colorants,pigments, stabilizers, preservatives, or antioxidants.

The pharmaceutical compositions can be formulated by techniques known tothe person skilled in the art, such as the techniques published inRemington's Pharmaceutical Sciences, 20th Edition. The pharmaceuticalcompositions can be formulated as dosage forms for oral, parenteral,such as intramuscular, intravenous, subcutaneous, intradermal,intraarterial, rectal, nasal, topical, aerosol or vaginaladministration. Dosage forms for oral administration include coated anduncoated tablets, soft gelatin capsules, hard gelatin capsules,lozenges, troches, solutions, emulsions, suspensions, syrups, elixirs,powders and granules for reconstitution, dispersible powders andgranules, medicated gums, chewing tablets and effervescent tablets.Dosage forms for parenteral administration include solutions, emulsions,suspensions, dispersions and powders and granules for reconstitution.Emulsions are a preferred dosage form for parenteral administration.Dosage forms for rectal and vaginal administration include suppositoriesand ovula. Dosage forms for nasal administration can be administered viainhalation and insufflation, for example by a metered inhaler. Dosageforms for topical administration include creams, gels, ointments,salves, patches and transdermal delivery systems.

The compounds according to the invention, in particular the compounds offormula 1, 2, 3, 4, 5 or 6, or the above described pharmaceuticalcompositions comprising one or more compounds of formula 1, 2, 3, 4, 5or 6 may be administered to a subject by any convenient route ofadministration, whether systemically/peripherally or at the site ofdesired action, including but not limited to one or more of: oral (e.g.as a tablet, capsule, or as an ingestible solution), topical (e.g.,transdermal, intranasal, ocular, buccal, and sublingual), parenteral (e.g., using injection techniques or infusion techniques, and including,for example, by injection, e.g. subcutaneous, intradermal,intramuscular, intravenous, intraarterial, intracardiac, intrathecal,intraspinal, intracapsular, subcapsular, intraorbital, intraperitoneal,intratracheal, subcuticular, intraarticular, subarachnoid, orintrasternal by, e.g., implant of a depot, for example, subcutaneouslyor intramuscularly), pulmonary (e.g., by inhalation or insufflationtherapy using, e.g., an aerosol, e.g. through mouth or nose),gastrointestinal, intrauterine, intraocular, subcutaneous, ophthalmic(including intravitreal or intracameral), rectal, and vaginal.

If said compounds or pharmaceutical compositions are administeredparenterally, then examples of such administration include one or moreof: intravenously, intraarterially, intraperitoneally, intrathecally,intraventricular, intraurethrally, intrastemally, intracranially,intramuscularly or subcutaneously administering the compoundspharmaceutical compositions, and/or by using infusion techniques. Forparenteral administration, the compounds are best used in the form of asterile aqueous solution which may contain other substances, forexample, enough salts or glucose to make the solution isotonic withblood. The aqueous solutions should be suitably buffered (preferably toa pH of from 3 to 9), if necessary. The preparation of suitableparenteral formulations under sterile conditions is readily accomplishedby standard pharmaceutical techniques well known to those skilled in theart.

Said compounds or pharmaceutical compositions can also be administeredorally in the form of tablets, capsules, ovules, elixirs, solutions orsuspensions, which may contain flavoring or coloring agents, forimmediate-, delayed-, modified-, sustained-, pulsed- orcontrolied-release applications.

The tablets may contain excipients, such as microcrystalline cellulose,lactose, sodium citrate, calcium carbonate, dibasic calcium phosphateand glycine, disintegrants, such as starch (preferably corn, potato ortapioca starch), sodium starch glycolate, croscarmellose sodium andcertain complex silicates, and granulation binders, such aspolyvinylpyrrolidone, hydroxypropylmethylcellulose (HPMC),hydroxypropylcellulose (HPC), sucrose, gelatin and acacia. Additionally,lubricating agents, such as magnesium stearate, stearic acid, glycerylbehenate and talc may be included. Solid compositions of a similar typemay also be employed as fillers in gelatin capsules. Preferredexcipients in this regard include lactose, starch, a cellulose, milksugar or high molecular weight polyethylene glycols. For aqueoussuspensions and/or elixirs, the agent may be combined with varioussweetening or flavoring agents, coloring matter or dyes, withemulsifying and/or suspending agents and with diluents, such as water,ethanol, propylene glycol and glycerin, and combinations thereof.

Alternatively, said compounds or pharmaceutical compositions can beadministered in the form of a suppository or pessary, or it may beapplied topically in the form of a gel, hydrogel, lotion, solution,cream, ointment or dusting powder. The compounds of the presentinvention may also be dermally or transdermally administered, forexample, by the use of a skin patch.

Said compounds or pharmaceutical compositions may also be administeredby the pulmonary route, rectal routes, or the ocular route. Forophthalmic use, they can be formulated as micronized suspensions inisotonic, pH adjusted, sterile saline, or, preferably, as solutions inisotonic, pH adjusted, sterile saline, optionally in combination with apreservative, such as a benzylalkonium chloride. Alternatively, they maybe formulated in an ointment, such as petrolatum.

For topical application to the skin, said compounds or pharmaceuticalcompositions can be formulated as a suitable ointment containing theactive compound suspended or dissolved in, for example, a mixture withone or more of the following: mineral oil, liquid petrolatum, whitepetrolatum, propylene glycol, emulsifying wax and water. Alternatively,they can be formulated as a suitable lotion or cream, suspended ordissolved in, for example, a mixture of one or more of the following:mineral oil, sorbitan monostearate, a polyethylene glycol, liquidparaffin, polysorbate 60, cetyl esters wax, 2-octyldodecanol, benzylalcohol and water.

Typically, a physician will determine the actual dosage which will bemost suitable for an individual subject. The specific dose level andfrequency of dosage for any particular individual subject may be variedand will depend upon a variety of factors including the activity of thespecific compound employed, the metabolic stability and length of actionof that compound, the age, body weight, general health, sex, diet, modeand time of administration, rate of excretion, drug combination, theseverity of the particular condition, and the individual subjectundergoing therapy.

A proposed, yet non-limiting dose of the compounds of formula 1, 2, 3,4, 5 or 6 for administration to a human (of approximately 70 kg bodyweight) may be 0.05 to 5000 mg, preferably 0.1 mg to 1000 mg, of theactive ingredient per unit dose. The unit dose may be administered, forexample, 1 to 4 times per day. The dose will depend on the route ofadministration. It will be appreciated that it may be necessary to makeroutine variations to the dosage depending on the age and weight of thepatient/subject as well as the severity of the condition to be treated.The precise dose and route of administration will ultimately be at thediscretion of the attendant physician or veterinarian.

The compounds of the present invention, including the compounds offormula 1, 2, 3, 4, 5 or 6, may be administered in the context of amonotherapy or in cotherapy with one or more other pharmaceuticalagents. For example, one compound of the present invention or two ormore compounds of the invention may be used in combination with one ormore immunomodulatory drugs and/or anti-inflammatory drugs for thetreatment, prevention or amelioration of an inflammatory, autoimmuneand/or allergic disorder.

A pharmaceutical composition may comprise said compound(s),immunomodulatory drug(s) and/or anti-inflammatory drug(s). Cotherapy mayalso include the administration of two or more compounds of the presentinvention in the absence of further immunomodulatory drugs oranti-inflammatory drugs. It is also envisaged herein that thecompound(s), immunomodulatory drug(s) and/or anti-inflammatory drug(s)might be linked, for example, by formation of conjugates. Accordingly,the compounds, immunomodulatory drugs and/or anti-inflammatory drugs maybe administered to a subject simultaneously. Also, a pharmaceuticalcomposition may comprise only the compound(s) of the present invention,while the one or more immunomodulatory drugs and/or anti-inflammatorydrugs are comprised in a different pharmaceutical composition. In thatcase, it may still be possible to administer the compound(s) of theinvention, immunomodulatory drugs and/or anti-inflammatory drugssimultaneously; however, the compound(s) of the invention may also beadministered before and/or after the one or more immunomodulatory drugsand/or anti-inflammatory drugs. It is readily apparent to a personskilled in the art how to administer, for example, one or more compoundsof the present invention, one or more immunomodulatory drugs, and/or oneor more anti-inflammatory drugs in cotherapy.

It is envisaged that one or more of the compounds as described herein,in particular the compounds of formula 1, 2, 3, 5 or 6, may be used incombination with one or more immunomodulatory drugs and/or one or moreanti-inflammatory drugs.

The one or more immunomodulatory drugs include, without being limitedthereto: antimetabolites such as, e.g., azathioprine, mycophenolic acid,leflunomide, teriflunomide, or methotrexate; macrolides such as, e.g.,tacrolimus, ciclosporin, or pimecrolimus; IL-2 inhibitors such as, e.g.,abetimus or gusperimus; TNF-α inhibitors such as, e.g., thalidomide orlenalidomide; IL-1 receptor antagonists such as, e.g., anakinra;mammalian target of rapamycin (mTOR) proteins such as, e.g., sirolimus,deforolimus, everolimus, temsirolimus, zotarolimus, or blolimus A9;monoclonal antibodies such as, e.g., eculizumab, infliximab, adalimumab,certolizumab pegol, afelimomab, golimumab, Mepolizumab, omalizumab,nerelimomab, faralimomab, elsilimomab, lebrikizumab, ustekinumab,muromonab-CD3,otelixizumab, teplizumab, visilizumab, clenoliximab,keliximab, zanolimumab, efalizumab, erlizumab, afutuzumab, ocrelizumab,pascolizumab, lumillximab, teneliximab, toralizumab, aselizumab,galiximab, gavilimomab, ruplizumab, belimumab, ipilimumab, tremelimumab,bertilimumab, lerdelimumab, metelimumab, natalizumab, tocilizumab,odulimomab, basiliximab, daclizumab, inolimomab, zolimomab aritox,atorolimumab, cedelizumab, dorlixizumab, fonlolizumab, gantenerumab,gomiliximab, maslimomab, morolimumab, pexelizumab, reslizumab,rovelizumab, siplizumab, talizumab, telimomab aritox, vapaliximab, orvepalimomab; polyclonal antibodies such as, e.g., anti-thymocyteglobulin or anti-lymphocyte globulin; or fusion proteins such as, e.g.,abatacept, belatacept, etanercept, pegsunercept, aflibercept, alefacept,or rilonacept.

Furthermore, the one or more anti-inflammatory drugs include, withoutbeing limited thereto: pyrazolidine or butylpyrazolidine derivativessuch as, e.g., ampyrone, clofezone, kebuzone, metamizole, mofebutazone,oxyphenbulazone, phenazone, phenylbutazone, sulfinpyrazone, orfeprazone; acetic acid derivatives such as, e.g., aceclofenac,acemetacin, alclofenac, bromfenac, bumadizone, bufexamac, diclofenac,difenpiramide, etodoiac, fentiazac, indometacin, ketorolac, lonazolac,oxametacin, proglumetacin, sulindac, tolmetin, zomepirac, or amfenac;oxicam derivatives such as, e.g., ampiroxicam, droxicam, lornoxicam,meloxicam, piroxicam, or lenoxicam; propionic acid derivatives such as,e.g., alminoprofen, benoxaprofen, dexibuprofen, dexketoprofen, fenbufen,fenoprofen, flunoxaprofen, flurbiprofen, ibuprofen, ibuproxam,indoprofen, ketoprofen, naproxen, oxaprozin, pirprofen, suprofen, ortiaprofenic acid; fenamic acid derivatives such as, e.g., flufenamicacid, meclofenamic acid, mefenamic acid, tolfenamic acid, niflumic acid,morniflumate, or azapropazone; COX-2 inhibitors such as, e.g.,celecoxib, etoricoxib, iumiracoxib, parecoxib, rofecoxib, or valdecoxib;or nabumetone, glucosamine, benzydamine, glycosaminoglycan, magnesiumsalicylate, proquazone, superoxide dismutase/orgotein, nimesulide,diacerein, tenidap, oxaceprol, or chondroitin sulfate.

Cotherapy using the compound(s) of the present invention,immunomodulatory drug(s) and/or anti-inflammatory drug(s) may result ina synergistic effect, i.e. the agents acting together may create aneffect greater than that predicted by knowing only the separate effectsof the individual agents. Such a synergistic effect might beparticularly advantageous if less amounts of the com pound (s),immunomodulatory drug(s) and/or anti-inflammatory drug(s) may then beused. Thus, possible side-effects of the compound(s), immunomodulatorydrug(s) and/or anti-inflammatory drug(s) might be diminished or avoided.

It is furthermore particularly envisaged that one or more of thecompounds of the invention, in particular the compounds of formula 1, 2,3, 4, 5 or 6, may be used in combination with one or moreimmunomodulatory drugs as described herein above and/or one or moreanti-inflammatory drugs as described herein above (including, forexample, azathioprine, ciclosporin, D-penicillamine, gold salts,hydroxychloroquine, leftunomide, methotrexate, minocycline,sulfasalazine, or cyclophosphamide) for the treatment, prevention oramelioration of rheumatoid arthritis.

The term “treatment of a disorder or disease” as used herein, such as“treatment of an inflammatory, autoimmune and/or allergic disorder”, iswell known in the art. “Treatment of a disorder or disease” implies thata disorder or disease is suspected or has been diagnosed in apatientysubject. A patient/subject suspected of suffering from adisorder or disease typically shows specific clinical and/orpathological symptoms which a skilled person can easily attribute to aspecific pathological condition (i.e., diagnose a disorder or disease).

“Treatment of a disorder or disease” may, for example, lead to a halt inthe progression of the disorder or disease (e.g., no deterioration ofsymptoms) or a delay in the progression of the disorder or disease (incase the halt in progression is of a transient nature only). “Treatmentof a disorder or disease” may also lead to a partial response (e.g.,amelioration of symptoms) or complete response (e.g., disappearance ofsymptoms) of the subject/patient suffering from the disorder or disease.“Amelioration” of a disorder or disease may, for example, lead to a haltin the progression of the disorder or disease or a delay in theprogression of the disorder or disease. Such a partial or completeresponse may be followed by a relapse. It is to be understood that asubject/patient may experience a broad range of responses to a treatment(e.g., the exemplary responses as described herein above).

Treatment of a disorder or disease may, inter alia, comprise curativetreatment (preferably leading to a complete response and eventually toheating of the disorder or disease) and palliative treatment (includingsymptomatic relief).

Also the term “prevention of a disorder or disease” as used herein, suchas “prevention of an inflammatory, autoimmune and/or allergic disorder”,is well known in the art. For example, a patient/subject suspected ofbeing prone to suffer from a disorder or disease as defined herein may,in particular, benefit from a prevention of the disorder or disease. Thesubject/patient may have a susceptibility or predisposition for adisorder or disease, including but not limited to hereditarypredisposition. Such a predisposition can be determined by standardassays, using, for example, genetic markers or phenotypic indicators, itis to be understood that a disorder or disease to be prevented inaccordance with the present invention has not been diagnosed or cannotbe diagnosed in the patient/subject (for example, the patient/subjectdoes not show any clinical or pathological symptoms). Thus, the term“prevention” comprises the use of compounds of the present inventionbefore any clinical and/or pathological symptoms are diagnosed ordetermined or can be diagnosed or determined by the attending physician.

The subject or patient, such as the subject in need of treatment,prevention or amelioration, may be a eukaryote, an animal, a vertebrateanimal, a mammal, a rodent (e.g. a guinea pig, a hamster, a rat, amouse), a murine (e.g. a mouse), a canine (e.g. a dog), a feline (e.g. acat), an equine (e.g. a horse), a primate, a simian (e.g. a monkey orape), a monkey (e.g. a marmoset, a baboon), an ape (e.g. gorilla,chimpanzee, orangutan, gibbon), or a human. The meaning of the terms“eukaryote”, “animal”, “mammal”, etc. is well known in the art and can,for example, be deduced from Wehner und Gehring (1995; Thieme Verlag).In the context of this invention, it is particularly envisaged thatanimals are to be treated which are economically, agronomically orscientifically important. Scientifically important organisms include,but are not limited to, mice, rats, rabbits, fruit flies like Drosophilamelagonaster and nematodes like Caenorhabditis elegans. Non-limitingexamples of agronomically important animals are sheep, cattle and pig,while, for example, cats and dogs may be considered as economicallyimportant animals. Preferably, the subject/patient is a mammal; morepreferably, the subject/patient is a human or a non-human mammal (suchas, e.g., a guinea pig, a hamster, a rat, a mouse, a rabbit, a dog, acat, a horse, a monkey, an ape, a marmoset, a baboon, a gorilla, achimpanzee, an orangutan, a gibbon, a sheep, cattle, or a pig; and inparticular a canine, such as a dog); even more preferably, thesubject/patient is a human. In this specification, a number of documentsincluding patent applications and manufacturer's manuals are cited. Thedisclosure of these documents, while not considered relevant for thepatentability of this invention, is herewith incorporated by referencein its entirety. More specifically, all referenced documents areincorporated by reference to the same extent as if each individualdocument was specifically and individually indicated to be incorporatedby reference.

The invention is also described by the following illustrative figures.The appended figures show: FIGS. 1A-1W: Inhibition of mast celldegranulation by compounds 1a (FIG. 1A), 1b (FIG. 1B), 1c (FIG. 1C), 1d(FIG. ID), 1e (FIG. 1E), 1f (FIG. 1F), 1g (FIG. 1G), 1h (FIG. 1H), 1i(FIG. 1I), 1j (FIG. 1J), 1k (FIG. IK), 1n (FIG. 1L), 1q (FIG. 1M), 1r(FIG. 1N), 1s (FIG. 1O), 1t (FIG. 1P), 1u (FIG. 1Q), 1v (FIG. 1R), 1w(FIG. IS), 1x (FIG. IT), 2a (FIG. 1U), 2b (FIG. 1V) and miltefosine(FIG. 1W). Dose-response curves for inhibition of p-hexosaminidaserelease from RBL-2H3 cells stimulated with antigen-specific lgE andtriggered with antigen are shown (means±standard error of the mean).

FIGS. 2A & 2B: Inhibition of Akt phosphorylation on Ser473 by compounds1a (FIG. 2A) and 1c (FIG. 2B). Percentage of total Akt phosphorylated onSer473 is expressed as a percentage of control untreated cells inducedwith IgE and antigen for 15 min (shown are means±standard deviation).

FIG. 3: Effect of compound 1a and dexamethasone on mouse ear swelling inthe DTH response in mice (data are means±standard deviations of 8mice; * p<0.01 vs. vehicle control (Dunnett's post hoc test)).

FIGS. 4A & 4B: Effect of compound 1a on mouse ear swelling in theallergic contact dermatitis model in mice. FIG. 4A shows the inhibitoryactivity of compound 1a at different administration times before antigenchallenge (data are means±SEM of 7 mice; * p<0.05 vs. vehicle control(Dunnett's post hoc test)). FIG. 4B shows the inhibitory activity ofcompound 1a after topical application (data are means±SEM of 7 mice; ***p<0.001 vs. vehicle control (Dunnett's post hoc test)).

The invention will now be described by reference to the followingexamples which are merely illustrative and are not to be construed as alimitation of the scope of the present invention.

EXAMPLES Example 1: 3-(N,N-Dimethylmyristylammonio)-Propanesulfonate 1a

Compound 1a is commercially available (Sigma-Aldrich Chemie GmbH,Munich, Germany, product number T7763).

Example 2: Preparation of 3-(N-Methyltetracylammonio)Propanesulfonate 1b

N-Methyltetradecylamine (454 mg, 2mmol) and 1,3-propanesultone (280 mg,2.3 mmol) are stirred in ethyl acetate (10 mL) for 24 h. The volatilesare removed and the residue is flash-chromatographed on silica usingdichloromethane/methanol (4:1). Rotary evaporation and drying in highvacuum yields 363 mg (52%) of 1b as a white solid.

¹H—NMR (300 MHz, CDCl₃) δ=0.81 (t. J-6.9, 3 H), 1.1-1.35 (m, 22H), 1.25(m, 2H), 2.22 (m, 2H), 2.81 (s, 3H), 2.98 (m, 4H), 3.25 (m, 2H).

MS (ESI): 350.3 (M+H⁺), 372.6 (M+Na⁺), 699.6 (2M+H⁺), 721.6 (2M+Na⁺).

Example 3: Preparation of 3—N,N-DimethylmyristylammoniopropylphosphonicAcid 1c

N,N-Dimethyltetradecylamine is alkylated with commercial diethyl3-bromopropyl-phosphonate to yield the corresponding ethyl ester, whichis purified by crystallization. Treatment with trimethylsilyibromide inthe presence of allyltrimethylsilana (Hammerschmidt 1991, Yan 2007)followed by hydrolysis of the resulting silylphosphonate yields compound1c as hydrobromide salt (1c—HBr).

The side products of the ester cleavage are volatile and can be removedin vacuum. 1c—HBr is subsequently purified by crystallization. Theamount of water used should be kept to a minimum because 1c and itshydrobromide tend to intense foaming during rotary evaporation.Attempted aqueous workup of the abovementioned ethyl ester, 1c—HBr or 1cresults in a stable emulsion. The hydrobromide is treated with exactlyone equivalent of NaOH, desalted by passing through a RP-18 column andthe resulting betaine 1c is purified by recrystallization.

¹ H—NMR (600 MHz, CDCl₃/CD₃OD 8:2): δ=0.90 (t, 3H), [1.22-1.43 (m), 1.39(br s), 1=22H)], 1.66 (d/t, J 17.3/7.0, 2H), 1.75 (br m, 2H), 2.02 (m,2H), 3.09 (s, 6H), 3.25 (m, 2H), 3.47 (m, 2H).

Example 4: Preparation of2-(N)N-Dimethyltetradecylammonio)Ethanesulfonate 1d

Sodium-2-bromoethansulfonate (411 mg, 1.95 mmol),N-methyltetradecylamine (342 mg, 1.50 mmol) and K₂CO₃ (269 mg, 1.95mmol) are suspended in dimethylformamide (DMF) (3 mL) and stirred atreflux overnight The volatiles are removed and the residue is purifiedby preparative HPLC to give, after drying at high vacuum, 324 mg of2-(N-methyltetradecylammonio)-ethanesulfonate as a white solid.

2-(N-Methyltetradecylammonio)-ethanesulfonate (113 mg, 0,25 mmol),methyl iodide (284 mg, 2.0 mmol) and K2CO3 (103 mg, 0.75 mmol) aresuspended in a mixture of acetone (3 mL) and dichloromethane (1 mL). Themixture is stirred at room temperature overnight, the solvent is removedand the residue purified by preparative HPLC to give 50 mg (57%) of 1d.

¹H—NMR (300 MHz, CDCl₃/CD₃OD 8:2); 5=0.80 (t, J=6.9, 3H), 1.1-1.45 (m,22H), 1.68 (m, 2H), 3.02 (s, 6H), 3.19 (m, 4H), 3.61 (m, 2H).

MS (ESI): 350.3 (M+H⁺), 699.6 (2M+H⁺).

Example 5: Preparation of 4-(N1N-Dimethylmyristylammonio)Butanesulfonate1e

1,4-Butane sullone (681 mg, 5 mmol) and N,N-dimethyltetradecylamin (966mg, 4 mmol) are dissolved in DMF (10 mL) and stirred under argonatmosphere at 130° C. bath temperature for 2 d. The volatiles areremoved and the residue is purified by preparative HPLC to yield 831 mg(55%) of 1e as white solid.

¹H—NMR (300 MHz, CDCl₃): δ=0.81 (t, J=6.9, 3H), 1.1-1.35(m, 22H), 1.61(m, 2H), 1.83 (m, 4H), 2.84 (m, 2H), 3.08 (s, 6H), 3.18 (m, 2H), 3.44(m, 2H). MS (ESI): 378.3 (M+H⁺). 755.6 (2M+H⁺).

Example 6: Preparation of2-((Dimethyl(Tetradecyl)Ammonio)Methyl)Butane-1-Sulfonate 1f

Ethylmalonic acid diethylester (1.86 g, 9.9 mMol) and N-methyltetradecylamine (1.5 g, 6.6 mmol) are dissolved in DMF (20 mL) and stirred at 130°C. bath temperature under argon atmosphere for 24 h. The mixture ispartitioned between water and EtOAc. The aqueous layer is extracted withEtOAc twice and the combined EtOAc layers are washed with sat. NaCl,dried over Na₂SO₄ and the solvent is removed. The residue is passedthrough a short plug of silica using dichloromethane/methanol 4:1, thesolvent removed under reduced pressure and the residue is dried in highvacuum to provide 2.1 g (86%) of ethyl2-(methyl(tetradecyl)carbamoyl)butanoate as a white material.

Under argon atmosphere, LiAlK, (608 mg, 16.0 mmol) is suspended in 10 mLof tetrahydrofuran (THF) and heated to reflux. A solution of ethyl2-(methyl(tetradecyl)-carbamoyl)butanoate (3.0 g, 8.0 mmol) in 15 mL ofTHF is added dropwise with caution and the resulting mixture is heatedto reflux overnight. Methanol is added dropwise with caution until theevolution of hydrogen ceases. 15 mL of water are added, whereupon thecolour changes from grey to white. The mixture is diluted with water andethyl acetate. The solids are filtered off using a pad of celite withEtOAc-washing. The aqueous layer is separated and extracted three timeswith ethyl acetate. The combined organic layers are washed with sat.NaCl (1x), dried over Na₂SO₄and the solvent is removed under reducedpressure. The residue is chromatographed on silica usingdichloromethane/methanol 10:1 to yield 550 mg (22%) of2-((methyl(tetradecyl)amino)methyl)butan-1-ol as a white solid.

2-((Methyl(tetradecyl)amino)methyl)butan-1-ol (390 mg, 1.24 mmol) isdissolved in dichloromethane (8 mL) under argon atmosphere.N,N-Diisopropylethylamine (DIEA) (163 mg, 1.30 mmol) and4-dimethylaminopyridine (DMAP) (15 mg, 0.12 mmol) are added. To thismixture is added dropwise methanesulfonyi chloride (148 mg, 1.30 mmol).The mixture is stirred at ambient temperature overnight The mixture isquenched with methanol and the solvent is removed under reducedpressure. Chromatography on silica using dichloromethane/methanol 10:1yields 208 mg (43%) of 2-((methyl(tetradecyl)amino)-methyl)butylmethanesulfonate.

2-((Methyl(tetradecyl)amino)-methyl)butyl methanesulfonate (200 mg, 0.5mmol) is dissolved in ethanol (2 mL). A solution of Na₂SO₃ (315 mg, 2.5mmol) in 1 mL of water is added and the mixture is stirred at 100° C.bath temperature for 3 h. After removal of the solvent under reducedpressure, the residue is chromalographed on silica usingdichloromethane/methanol 20:1 to yield 120 mg (64%) of2-((methyl(tetradecyl)ammonio)methyl)butane-1-sulfonate.2-((Methyl(tetradecyl)ammonio)methyi)butane-1-sulfonate (90 mg, 0.23mmol) is dissolved in 2 mL of dichloromethane under argon atmosphere.K₂CO₃ (97 mg, 0.70 mmol) and methyl iodide (255 mg, 1.80 mmol) are addedand the mixture is stirred at room temperature overnight. After removingthe solvent under reduced pressure, the residue is purified bypreparative HPLC to yield 78 mg (87%) of 1 f as a white solid.

¹H—NMR (300 MHz, CDCl₃): 5=0.81 (t, J=6.9, 3H), 0.92 (t, J=7.3, 3H),1.1-1.35 (m, 22K), 1.50 (m, 2H), 1.66 (m, 2H), 2.40 (m, 1H), 2.75 (m,1H), 2.91 (d/m, J=12.5, 1H). 3.10 (m, 1H), 3.12/3.16 (2s, 3H), 3.26 (m,2H), 4.06 (d/m, J=12.9,1H).

MS (ESI): 392.4 (M+H⁺), 783.7 (2M+H⁺).

Examples 7, 8 and 9: Preparation of3-(N,N-Dimethyloctadecylammonio)Propanesulfonate 1g,3-(N,N-dimethyIpalmitylammonio)propansulfonate 1h and3-(N,N-dimethyldodecyl-ammonio) propanesulfonate 1i

Compounds 1g, 1h and 1i are prepared in a similar way as described forcompound 1b using N-methyloctadecylamine (for 1g),N-methylhexadecylamine (for 1h) or N-methyldodecyl-amine (for 1i)instead of N-methyltetradecylamine followed by quarternization of thenitrogen using methyl iodide as described in the final preparation stepfor compound 1d.

Alternatively, compounds 1g, 1h and 1i are commercially available fromSigma-Aldrich GmbH, Munich, Germany (1g: product number 41570; 1h:product number H6883; 1i: product number D0431).

Example 10: N-Tetradecyl—N,N-Dimethylglycine 1j

Compound 1j is commercially available (Affymetrix, Santa Clara, Calif.95051, USA, product number T305).

Example 11: Preparation of 3-(Dimethyl(Tetradecyl)Ammonio)Propanoate 1k

Under argon atmosphere, (5-propiolactone (216 mg, 3.0 mmol) is dissolvedin a mixture of 4 mL of ether and 2 mL of acetonitrile.N,N-dimethyltetradecylamine (724 mg, 3.0 mmol) is added dropwise over aperiod of 2 h. The mixture is stirred for another 30 min and theproduct, a white precipitate, is collected by filtration, washed withseveral portions of ether and dried in vacuo, yielding 470 mg (50%) of1k as a white powder. The product is stored below −15° C.

¹H—NMR (300 MHz, CDCl₃): 8=0.81 (t, J=6.4, 3H). 1.1-1.35 (m, 22H), 1.65(m, 2H), 2.55 (t. J=7.8, 2H), 3.13 (s, 6H), 3.20 (m, 2H), 3.69 (t,J=7.5, 2H). MS (ESI): 314.3 (M+H⁺), 627.5 (2M+H⁺).

Example 12: Preparation of 3-(Dimethyl(Dodecyl)Ammonio)Propanoate 1m

Under argon atmosphere, (3-propiolactone (216 mg, 3.0 mmol) is dissolvedin a mixture of 4 mL of ether and 2 mL of acetonitrile.N,N-dimelhyldodecylamine (639 mg, 3.0 mmol) is added dropwise over aperiod of 2 h. The mixture is stirred overnight and the product, a whiteprecipitate, is collected by filtration, washed with several portions ofether and dried in vacuo, yielding 653 mg (76%) of 1m as a white powder.The product is stored below −15° C. ¹H—NMR (300 MHz, CDCl₃): 6=0.81 (t,J=6.4. 3H). 1.1-1.35 (m, 22H), 1.65 (m, 2H), 2.55 (t, J=7.8, 2H). 3.13(s, 6H), 3.20 (m, 2H), 3.69 (t, J=7.5, 2H). MS (ESI): 314.3 (M+H⁺),627.5 (2M+H⁺).

Example 13: Preparation of 4-(Methyl(Tetradecyl)Ammonio)Butanoate 1n

To a solution of 681 mg (3.0 mmol) of N-methyl tetradecylamine inabsolute dimelhylformamide (DMF) (6 mL), are added 834 mg (6.0 mmol) ofpowdered K₂CO₃ and 1.34 g (6.0 mmol) of 4-bromobutyric acid. The mixtureis stirred under argon atmosphere at 130° C. bath temperature for 3 d,The votatiles are removed on a rotary evaporator and the residuepurified by preparative HPLC to yield 594 mg (41%) of4-(methyl(tetradecyl)ammonio)butyric acid tert-butyl ester as a TFAsalt. ESI-MS (pos.): 370.3 (M+H⁺).

4-(methyl(tetradecyl)ammonio)butyric acid tert-butyf ester (241 mg, 0.5mmol) is suspended in 3 mL of TFA/H2O (95:5). The mixture is stirred for1 h at room temperature, the volatiles removed on a rotary evaporatorand the residue purified by preparative HPLC to yield 114 mg (36%) of 1n as a white solid.

¹ H—NMR (300 MHz, CDCl₃/CD₃ 8:2): 5=0.73 (t, J=6.9, 3H), 1.11 (m), 1.18(m, X=22H), 1.56 (m, 2H), 1.85 (m, 2H), 2.29 (t, J=6.7. 2H), 2.66 (s,3H). 2.89 (m), 2.97 (m. X=4H), 4.05 (br. s, 3H).

MS (ESI): 314.2 (M+H⁺), 627.4 (2M+H⁺). Neg.: 312.0 (M—H⁻).

Example 14: Preparation of 4-(Dimethyl(Tetradecyl)Ammonio)Butanoate 1o

Under argon atmosphere, 4-(methyl(tetradecyl)ammonio)butyric acidtert-butyl ester, TFA salt (described for 1n) (338 mg, 0.70 mmol) isdissolved in acetone (5 ml). Dry powdered K₂CO₃ (486 mg. 3.5 mmol) andmethyl iodide (497 mg, 3.5 mmol) are added and the mixture stirredovernight. The product is purified by preparative HPLC and dried in highvacuum, yielding 266 mg (53%) of 4-(dimethyl(tetradecyl)ammonio)butyricacid tert-butyl ester, TFA salt, 100% pure by HPLC. This material isstirred with 3 ml of TFA/H20 (95:5) for 1 h. HPLC indicated completeconversion. The volatiles are removed on a rotary evaporator and theresidue purified by preparative HPLC and dried in high vacuum to yield105 mg (60%) of 1o.

¹H—NMR (300 MHz, CDCl₃): 6=0.86 (t, J=6.4. 3H), 1.15-1.35 (m, 22H), 1.69(m, 2H). 1.98(m, 2H), 2.41 (t, J=6.1, 2H). 3.06 (s, 6H), 3.17 (m, 2H),3.34 (m, 2H).

MS (ESI): 328.3 (M+H⁺), 655.6 (2M+H⁺).

Example 15: N-Dodecyl—N,N-Dimethylglycine 1p

Compound 1p is commercially available (Affymetrix, Santa Clara, Calif.95051, USA, product number D350).

Example 16: Preparation of 2-(N-Methyldodecylammonio)Ethanesulfonate 1q

Sodium-2-bromoethane sulfonate (411 mg, 1.95 mmol) and powdered K₂CO₃(269 mg, 1.95 mmol) are suspended in 3 mL of dry DMF under argonatmosphere. N-methyldodecylamine (298 mg, 1.50 mmol) is added and themixture stirred at 130° C. overnight. The volatiles are removed on arolary evaporator and the residue purified by preparative HPLC and driedin high vacuum to yield 342 mg (74%) of 1q as a white solid.

¹ H—NMR (300 MHz, CDCl₃); 5=0.81 (t, J=6-4, 3H), 1.10-1.35 (m, 18H),1.70 (m, 2H)f 2.88 (s), 2.89 (s, Σ=3H), 3.0-3.45 (m, 5H), 3.55 (m, 1H),8.46 (br. s, 1H), 8.89 (br. s, 1H). MS (ESI): 308.2 (M+H⁺), 325.3 (M+NH₄⁻), 615.4 (2M+H⁺).

Example 17: Preparation of 2-(N,N-Dimethyldodecylammonio)Ethanesulfonate1r

A mixture of 2-(N-methyldodecylammonio)ethanesulfonate 1q (225 mg, 0.73mmol), methyl iodide (568 mg, 4.0 mmol), potassium carbonate (207 mg, 15mmol) and acetone (5 mL) is stirred under argon atmosphere for 2.5 d.The volatiles are removed and the residue is purified by preparativeHPLC and dried in high vacuum to yield 161 mg (68%) of product 1r.

¹H—NMR (300 MHz, CDCl₃/CD₃OD 8:2): δ=0.80 (t, J=6.5, 3H), 1.18/1.29 (2m,Σ=18H), 1.68 (m, 2H), 3.02 (s, 6H), 3.20 (m, 4H), 3.61 (m, 2H).

MS (ESI): 322.2 (M+H⁺), 643.5 (2M+H⁺), 660.5 (2M+NH₄ ⁺, 665.5 (2M+Na⁺).

Example 18: Preparation of 2-(N-Methyltetradecylammonio)Ethanesulfonate1s

A mixture of sodium-2-bromoethanesulfonate (411 mg, 1.95 mmol),N-methyltetradecylamine (342 mg, 1.50 mmol), K2C03 (269 mg, 1.95 mmol)and DMF (3 mL) is stirred at 135° C. bath temperature under argonatmosphere overnight. The volatiles are removed on a rotary evaporatorand the residue purified by preparative HPLC to yield 324 mg (64%) of1s. ¹H—NMR (300 MHz, CDCl₃): 5=0.90 (t, J=6.4, 3H), 1.15-1.45 (m, 22H),1.78 (m, 2H), 2.97/ 2.99 (2s, Σ=3H), 3.05-3.55 (m, 5H), 3.64 (m, 1H),7.36 (br. s, 1H). MS (ESI): 336.3 (M+H⁺), 671.5 (2M+H⁺), 693.5 (2M+Na⁺),709.5 (2M+K⁺).

Example 19: Preparation of3-(N,N-Dimethyldodecylammonio)Propylphosphonic Acid 1t

Under argon atmosphere, diethyl(3-bromopropyl)phosphonate (1.50 g. 5.75mmol) is dissolved in 5 mL of absolute ether. N,N-dimethyldodecylamine(1.07 g, 5.00 mmol) is added and the mixture stirred overnight. Thevolatiles are removed and the residue dried in vacuo, resulting insolidification. The hygroscopic solid is broken up and triturated withether and the ether removed by suction filtration. The residue is driedin high vacuum to yield 1.46 g (62%) of3-(N,N-dimethyldodecylammonio)propylphosphonic acid diethylester(bromide salt) as a white, hygroscopic solid.

¹H—NMR (300 MHz, CDCl₃): 6=0.81 (t, J=6.4, 3H), 1.19 (m), 1.27 (t,J=7.05, I=24H), 1.67 (m, 2H), 1.81 (d/t, J=18.1/6.8, 2H), 2.00 (m, 2H),3.36 (s, 6H), 3.42 (m, 2H), 3.73 (m, 2H), 4.04 (m, 4H). MS (ESI): 392.4(M+H⁺). 3-(N,N-dimethyldodecylammonio)propylphosphonic aciddiethylester, bromide salt (175 mg, 0,37 mmol) is placed under argonatmosphere, Absolute dichloromethane (3 mL), bromotrimethylsilane (233iL, 1.8 mmol) and allyltrimethylsilane (143 pL, 0.9 mmol) are added andthe mixture stirred at room temperature for 2.5 d. The volatiles areremoved on a rotary evaporator and the residue purified by preparativeHPLC to yield 106 mg (85%) of 1t. 1H—NMR (300 MHz, CDCl₃/CDCl₃ 8:2):5=0.78 (t, J=6.3, 3H), 1.1-1.3 (m, 18H), 1.62 (m, 4H), 2.10 (m, 2H),2.97 (s, 6H), 3.13 (m, 2H), 3.27 (m, 2H). MS (ESI): 336.2 (M+H⁺), 671.5(2M+H⁺).

Example 20: Preparation of 3-(N-Methyl—N—Hexadecylamino)PropylphosphonicAcid Diethylester Hydrobromide 1w

Under argon atmosphere, N-methylhexadecylamine (510 mg, 2.0 mmol) ispartially dissolved in absolute ether (4 mL).Diethyl(3-bromopropyl)phosphonate (647 mg, 2.50 mmol) anddiisopropylethylamine (436 jjL, 2.5 mmol) are added and the mixturestirred overnight at room temperature. The volatiles are removed and theresidue purified by flash chromatography on silica usingdichloromethane/methanol 10:1 to yield 216 mg (21%) of 1w.

¹H—NMR (300 MHz, CDCl₃): 8=0.81 (t, J=6.3, 3H), 1.19 (m), 1.25 T, J=6.9,X=32H), 1.40(m, 2H), 1.72 (m, 4H), 2.18 (s, 3H), 2.31 (m, 2H), 2.39 (br.t, J=6.5. 2H), 4.02 (mc, 4H). MS (ESI): 434.4 (M+H⁺). 889.7 (2M+Na⁺).

Example 21: Preparation of 3-(N-Methyl—N—Hexadecylamino)PropylphosphonicAcid 1v

Under argon atmosphere, a mixture of3-(N-methyl—N-hexadecylamino)propylphosphonic acid diethyiesterhydrobromide (1w) (130 mg, 0.25 mmol), absolute dichloromethane (2 mL),bromotrimethylsiiane (184 mg, 1.20 mmol) and allyltrimethylsilane (68mg, 0.60 mmol) is stirred overnight at room temperature. The volatilesare removed and the residue purified by preparative HPLC to yield 52 mg(55%) of 1v.

¹H—NMR (300 MHz, CDCl₃/CD₃OD 8:2):=0.89 (t, J=6.5, 3H), 1.2-1.4 (m,26H), 1.65-1.87 (m, 4H), 2.06 (m, 2H), 2.80 (s, 3H), 2.85-3.25 (br. m,4H). MS (ESI): 378.3 (M+H⁺), 755.6 (2M+H⁺).

Example 22: Preparation of3-(N-Methyl-N-Tetradecylamino)Propylphosphonic Acid DiethyiesterHydrobromide 1x

N-methyltetradecylamine (454 mg, 2.0 mmol) is dissolved in absoluteether (3 mL) under argon atmosphere. Diispropylethylamine (322 mg, 2.5mmol) and diethyl(3-bromopropyl) phosphonate (647 mg, 2.5 mmol) areadded and the mixture stirred for 2.5 d at room temperature. Aprecipitate Is removed and the supernatant concentrated on a rotaryevaporator and purified by flash chromatography on silica usingdichoromethaneAnethanol (10:1) to yield 356 mg (26%) of 1 x a whitesolid.

¹ H—NMR (300 MHz, CDCl₃): 5=0.81 (t, J=6.9, 3H), 1.19 (m), 1.25 (t,J=7.1, 2=28H), 1.40 (m, 2H), 1.71 (m, 4H), 2.18 (s, 3H), 2.31 (m, 2H),2.39 (br. t, J=7.2, 2H), 2.95 (br. s, 1H), 4.02 (mc, 4H). MS (ESI):406.4 (M+H⁺), 833.6 (2M+Na⁺).

Example 23: Preparation of3-(N-Methyl—N-Tetradecylamino)Propylphosphonic Acid (Betaine) 1u

3-(N-methyl—N-tetradecylamino)propylphosphonic acid diethyiesterhydrobromide 1x (162 mg, 0.33 mmol) is dissolved in 2 mL of absolutedichloromethane under argon atmosphere. Trimethylsilyl bromide (245 mg,1.6 mmol) and allyltrimethylsilane (91 mg, 0.8 mmol) are added and themixture stirred overnight at room temperature. The volatiles are removedon a rotary evaporator and the residue dissolved in ethanol and purifiedby preparative HPLC to yield 105 mg (91%) of 1u as a white solid. 1H—NMR(300 MHz, CDCI3): 5=0.81 (t, J=6.9, 3H), 1.1-1.3 (m, 20H), 1,61 (br.s,2H), 1.75 (m, 2H), 2.00 (m, 2H), 2.73 (s, 3H), 2.86 (m), 3.02 (m). 3.14(m, X=4H). MS (ESI): 350.3 (M+H⁺)r 699.6 (2M+H⁺). Neg.: 697.3 (2M—H⁺).

Examples 24 and 25: Preparation ofN—Hexadecyl—N-(3-Sulfonatopropyl)Piperidinium 2a and1—Hexadecyl-1-(3-Sulfonatopropyl)-4—Hydroxypiperidinium 2b

Piperidine (1.32 g, 15.5 mmol) and 1-iodohexadecane (1.82 g, 5.17 mmol)are dissolved in 6-8 mL of ethanol and stirred overnight while shieldedfrom light. The mixture is partitioned between plenty of ether anddilute NaOH, washed with sat. NaCl once, dried over Na₂SO₄, rotavappeddown to an oil and dried in vacuo. Yield: 1.45 g (90% of brownish oil).The product is used in the next step without further purification.

In 4 mL of EtOAc, N-hexadecylpiperidine (300 mg, 0.97 mmol) is mixedwith 1,3-propanesultone (142 mg, 1.63 mmol). The mixture is stirredfirst at room temperature (2 d), then at 50° C. overnight. Afteraddition of another 142 mg of 1,3-propanesultone, the mixture is stirredat reflux for 2 days, at which point analytical HPLC indicates completeconversion. The volatiles are removed and the residue is purified bypreparative HPLC to yield 298 mg (71%) of pure 2 a as a white solid.

¹ H—NMR (300 MHz, CDCl ₃): 5=0.81 (t, J=7.8, 3 H). 1,1-1.4 (m, 26 H),1.5-1.8 (m, 6 H), 1.91 (m, 2 H), 2.14 (m, 2 H), 2.91 (t, J=6.2, 2H),3.16 (m, 2H), 3.29 (m, 2H), 3.45 (M, 2H), 3.64 (m, 2H).

MS (ESI): 432.4 (M+H⁺), 863.8 (2M+H⁺).

Compound 2b is prepared in a similar way using 4-hydroxypiperidineinstead of piperidine.

Example 26: Preparation ofTrans—N-Tetradecyl—N-(3-Sulfonatopropyl)-3-Diethylaminocarbonyl-Piperidinium2c

A mixture of N,N-diethyl-3-piperidinecarboxamide (809 mg, 4.4 mmol).ethanol (3 mL) and tetradecyl iodide (620 mg, 1.9 mmol) is stirredovernight at room temperature. HPLC shows complete conversion. Themixture is diluted with ether (200 mL) and extracted with 1N NaOH (1x100 mL) and saturated NaCl (1x 100 mL) and the organic layer dried overNa₂SO₄ and filtered. The volatiles are removed and the residue purifiedby flash chromatography on silica using dichloromethane/methanol (10:1).The product is dried in vacuo to yield 682 mg (94%) of1-tetradecyl-3-dielhylaminocarbonylpiperidine. The product containsresidual N,N-diethyl-3-piperidinecarboxamide, which is removed in thenext step.

MS (ESI): 381.4 (M+H⁺).

A mixture of 1-tetradecyl-3-diethylaminocarbonylpiperidine (380 mg, 1.0mmol), ethyl acetate (3 mL) and 1,3-propanesultone (488 mg, 4.0 mmol) isstirred under argon atmosphere under reflux for 6 d. HPLC showsconversion is incomplete and the cis- and trans-stereoisomers give riseto two closely eluting peaks. The volatiles are removed and the residuepurified by preparative HPLC. The two resulting fractions are purifiedagain by preparative HPLC, yielding the racemic cis- andtrans-stereoisomer in 95% purity, with a yield of 71 mg (14%) oftrans-stereoisomer 2c and 54 mg (11%) of the cis-stereoisorner.

¹ H—NMR (125 MHz, CDCI3): 5=0.81 (t, J=6.4, 3H), 1.03 (t, J=7.1, 3H),1.10-1.35 (m, 25H), 1.50 (m, 1H), 1.60-1.90 (m, 3H), 1,90-2.20 (m, 3H),2.20-2.45 (m, 1H), 2.85-3.15 (m, 5H), 3.15-3.55 (m, 7H), 3.60 (br. d,J=12.2,1H), 3.75-3.95 (m, 3H). ,3C—NMR and DEPT (125.7 MHz, CDCl3):12.73 (CH3), 14.09 (CH3), 14.74 (CH3), 18.15 (CH2), 18.88 (CH2), 21.60(CH2), 22.66 (CHZ), 25.91 (CH2), 26.37 (CH2), 28.99 (CH2), 29.32 (CH2),29.38 (CH2), 29.52 (CH2), 29.60 (CH2), 29.62 (CH2), 31.88 (CH2), 32.92(CH), 40.51 (CH2), 40.51 (CH), 42.15 (CH2), 47.09 (CH2), 53.80 (CH2),59.20 (CH2), 60.64 (CH2). 65.23 (CH2), 170.09 (CO). MS (ESI): 503.5(M+H⁺).

Example 27: Inhibition of Mast Cell Degranulation Introduction

Mast cells are key effector cells involved in allergic and inflammatorydiseases, and the Rat Basophilic Leukemia clone 2H3 (RBL-2H3) cell lineis a commonly used model of allergen dependent immune modulator release(degranulation) in mast cells. On their surface, they express the highaffinity receptor for IgE (FceRI). Upon binding of antigen-specific IgEto the receptor, cells become sensitized to the IgE specific antigen(allergen). When IgE-sensitized cells then encounter multivalentantigen, the antigen clusters IgE-FceRI complexes and initiates a signaltransduction cascade that leads to degranulation, that is, the releaseof inflammatory mediators, such as cytokines, eicosanoids, histamine andenzymes. The assay can be used as a screening method to identifyimmune-modulating compounds, in particular compounds useful in themedical management of allergic and inflammatory diseases and asthma,β-hexosaminidase was previously shown to be released with the samekinetics as histamine (Schwartz et al., J Immunology; 123:1445-1450(1979)), thus offering a simple means to monitor degranulation. TheRBL-2H3 cell line has been successfully used to identify compounds withanti-allergic activity (Choo et al. Planta Med., 69:518-522 (2003)).

Materials and Methods

Materials Chemicals: Rat anti-DNP IgE monoclonal antibody was acquiredfrom Biozol (BZL06936), dinitrophenyl-conjugated human serum albumin(A6661) and Triton X-100 (T9284) were from Sigma-Aldrich,4-methylumbelliferyl—N-acetyl-β-D-glucosaminide (474502),Phorbol-12-myristate-13-acetate (524400) and thapslgargin (586005) fromCalbiochem. lonomycin (ALX-450-006) was purchased from AlexisBiochemicals. DMSO was from Merck (1.02950.0500) or Sigma-Aldrich(D2650). Cell culture media and supplements, Minimum Essential Medium(21090-022), Minimum Essential Medium without Phenol Red (51200-046),RPMI 1640 Medium (31870-025), L-Glutamine (25030-24) and 0.05%Trypsin-EDTA (25300-054), were obtained from Invitrogen. Fetal bovineserum (A15-151) was from PAA Laboratories. Other reagents were standardlaboratory grade or better.

Buffers and solutions: Phosphate buffered saline (PBS) and 1 M HEPESwere provided by the in-house service facility. Tyrode's buffer (TyB)consisted of Minimum Essential Medium without Phenol Red supplementedwith 2 mM L-gtutamine and 20 mM HEPES. Lysis buffer consisted of 25 mMTris—HCl, pH 7.5, 150 mM NaCl, 5 mM EDTA and 0.1% (w/v) Triton X-100.DNP—HSA was dissolved to 1 mg/ml in water. MUG substrate solutionconsisted of 2.5 mM 4-methylumbelliferyl—N-acetyl-β-D-glucosaminide in0.05 M citrate, pH 4.5; stop solution was 0.1 M NaHCO₃/0.1 M Na₂CO₃, pH10.

Consumables and equipment: For small-volume liquid handling procedures,Rainin LTS electronic pipettes were routinely used (Mettler-Toledo).Costar-Corning 24-well plates (3337) were centrifuged in an Eppendorf5804 R centrifuge. A Heraeus B15 table top incubator was used forincubations at 37° C. under non-sterile conditions. Fluorescence wasmeasured in black Nunc 96-well plates (237105) using a microplate reader(Tecan Safire) or FlexStation 3 (Molecular Devices) multi-mode platereader. Cells were maintained in Hera Cell 240CO₂ incubators (ThermoScientific). Serological pipettes (4487, 4488 and 4489) and cell cultureflasks (431080) were from Coming-Co star, 1.5 and 2 ml microcentrifugetubes (0030 120.086 and 0030 120.094) were from Eppendorf.

Cell Culture: RBL-2H3 cells obtained from the German Collection ofMicroorganisms and Cell Cultures (ACC312) (Braunschweig, Germany) weremaintained in 70% Minimum Essential Medium with Earle's Salts, 20% RPMl1640 Medium, 10% FBS and 2 mM l-glutamine in 95% air/5% CO₂ at 37° C androutinely checked for mycoplasma contamination. Cells were passagedevery 3-4 days; after washing cells once with 35 ml PBS cells wereincubated 8 min with 5 ml 0.05% Trypsin-EDTA solution at 37° C. Cellswere removed from the incubator, 15 ml culture medium was added andcells were resuspended by repeated pipetting.

Cell seeding: cells were harvested with Trypsin-EDTA as described and50-100 μl cell suspension seeded Into Costar CellBind 24 well clusterplates (no. 3337). Plates were kept for 30 min at RT under the sterilehood before being transferred to the incubator. Cells were used withinone or two days after seeding.

Measurement of β—Hexosaminidase Release Experimental Procedures

For sensitization, cells for immediate use were sensitized 6-12 h afterplating; cells to be used the following day were sensitized 26-38 hafter plating. Culture plates were removed from the incubator andchecked for cell growth and contamination. The medium was discarded andcells were sensitized with anti-DNP IgE (0.4 μg/ml) in 0.4 ml culturemedium overnight. Following overnight sensitization, cells were washedwith 0.8 ml pre-warmed TyB and 0.38 ml test compound or vehicle control(supplemented or not with 1% FBS) were added to duplicate wells. Sampleswere adjusted to contain 1% vehicle for test compounds dissolved inorganic solvents. Cells were incubated for 1 h at 37° C. At the end ofthe incubation period, cells were routinely stimulated with 20 μlDNP-HSA (2 μg/ml; final concentration 0.1 μg/ml) diluted in TyB andcells were incubated for 15 min at 37° C. Alternatively, cells werestimulated with 20 μl μM ionomycin (final concentration 0.25 μM) or 20nM PMA thapsigargin (final concentration 0.25 μM), both in the absenceor presence of 20 nM PMA (final concentration).

Plates were removed from the incubator and immediately centrifuged at 4°C. for 5 min at 250×g and transferred to an ice bath. Aliquots ofsupernatants, 25 μl, were transferred to 96-well plates. Remainingsupernatant was aspirated from control wells and cells were lysed in 400μm lysis buffer for 5 min at RT on an orbital shaker at 450 rpm undernon-sterile conditions. After lysis, 25 μl aliquots of lysates weretransferred to 96-well plates. MUG substrate solution, 100 μl, wereadded to supernatant and lysate samples and plates were incubated 30 minat 37° C. The reaction was terminated by addition of 150 μl stopsolution. Fluorescence was measured at 365 nm excitation and 440 nmemission wavelengths.

Test compound preparation: test compounds were prepared in 1.5 or 2 mlmicrocentrifuge tubes and incubated for 30 min at 37° C in a ThermomixerComfort with agitation (750 rpm). An electronic multichannel pipette wasused for rapid transfer of compound dilutions from microcentrifuge tubesto the cells.

Controls: conlrols used are defined as follows: negative control,supernatant of unstimulated cells was measured for unspecificβ-hexosaminidase release; positive control, supernatant of DNP-HSAstimulated cells was measured for specific, antigen-stimulatedβ-hexosaminidase release; maximum control, lysate of unstimulated cellswas measured for total p-hexosaminidase content.

Assessment of Pharmacologic Effect

Degranulation (β-hexosaminidase release): Degranulation was calculatedas the percentage of β-hexosaminidase released with respect to maximumcontrol (total β-hexosaminidase) after subtraction of negative control(unspecific release) using the formula;

% Degranulation=100 * 1—(test compound-negative control)/(maximumcontrol-negative control)

% Inhibition of degranulation (inhibition of β-hexosaminidase release):Inhibition of degranulation was calculated as percent reduction ofβ-hexosaminidase release with respect to positive control(antigen-stimulated release) after subtraction of negative control(unspecific release) using the formula;

% Inhibition=100 * (1—(test compound-negative control)/(positivecontrol-negative control))

Measurement of maximum tolerated concentration

The maximum tolerated concentration (MTC), i.e. the highestconcentration of test compound that does not cause cytotoxicity, asdetermined by the release of lactate dehydrogenase, was measured overthe tested concentration range. A commercially available cytotoxicitytest was used (Promega Cytotox-One cat. #67891).

The safety index (SI) of a test compound Is the ratio between themaximum tolerated concentration and the 1C50 and is used as a measure ofthe relative safety of the test compound.

Results

Concentration-dependent inhibition of degranulation was determined forail test compounds over a concentration range, as shown in FIG. 1, andIC50 values (concentration at which 50% of maximal inhibition isreached) were determined for each compound together with the MTC valuesover the same concentration range (Table 1). Results are taken from atleast three independent experiments.

TABLE 1 Inhibition of degranulation: IC50, MTC and SI values CompoundIC50 (μM) MTC (μM) SI 1a 3.2 100 31.3 1b 4.3 100 23.3 1c 2.8 200 71.4 1d3.9 100 25.6 1e 3.7 150 40.5 1f 4.0 75 18.8 1g 9.5 50  5.3 1h 3.7 5013.5 1i 4.1 200 48.8 1j 5.1 75 14.7 1k 5.3 100 18.9 1m 73% 100 —inhibition at 25 μM 1n 8.0 100 12.5 1o 65% 100 — inhibition at 25 μM 1p50% 100 — inhibition at 25 μM 1q 8.8 200 22.7 1r 4.2 200 47.6 1s 5.6 10017.9 1t 6.1 200 32.8 1u 5.9 200 33.9 1v 8.8 200 22.7 1w 6.9 200 29.0 1x5.5 100 18.2 2a 4.5 50 11.1 2b 4.3 50 11.6 2c 50% 100 — inhibition at 25μM Miltefosine 4.2 25  6.0

The MTC of the test compounds was 5-70 fold higher than their respectiveIC50s and hence, the inhibition of degranulation can be ascribed to apharmacological effect and not to an effect secondary to cytotoxicity.

All substances outlined in Table 1 show IC50 values in the lowmicromoJar range combined with high MTC values when compared toMiltefosine. Thus, the compounds according to the invention and, inparticular compounds 1a to 1x and compounds 2a to 2c, have anadvantageously low cytotoxicity.

Mast cell degranulation is a key cellular event in allergic andinflammatory reactions, in particular in pathological events involvingthe release of mediators such as histamine, leukotrienes andprostaglandins as well as proteases. As consequence, the inhibition ofmast cell degranulation is a valuable strategy for prevention ortreatment of pathological processes involving the aforementionedmediators. Furthermore, the mast cell degranulation assay provides anestimate of the activity of test compounds in other cells that play akey role in the inflammatory response, such as granulocytes, macrophagesand thymocytes, which release proinflammatory cytokines and chemokinesand tissue eroding proteases.

Example 28: Inhibition of Activation of Akt Kinase Introduction

The mast cell degranulation assay using the RBL-2H3 cell line (seeExample 27) was also used to determine the status of the PI3K/Akt axis.Activation of PI3K leads to production of PIP3 on the cytosolic side ofthe lipid bilayer. Akt is recruited to the PIP3 domain and subsequentlyactivated by phosphorylation on residues Ser473 and Thr308. (Franke etal., Cell 81:727-736, (1995)). Once recruited to the membrane, it isphosphorylated and activated by other kinases (Hemmings, Science275:628-630 (1997); Hemmings. Science 276:534 (1997); Downward, Science279:673-674 (1998); Alessi et al., EMBO J. 15:6541- 6551 (1996)).Western blotting of the phosphorylated Ser473 residue on Akt(phospho-Akt Ser473) is widely used to assess the level of activation ofthe PI3K/Akt axis.

Materials and Methods Materials

AH buffers and solutions used for the phosphor-Akt Ser473 assay werefrom Meso Scale Discovery. Tris Lysis Buffer consisted of 150 mM NaCl,20 mM Tris, pH 7.5, 1 mM EDTA, 1 mM EGTA and 1% Triton-X-100. CompleteTris Lysis Buffer was prepared prior to use by addition of proteaseinhibitor, phosphatase inhibitors and PMSF. The 10 x Tris Wash Bufferconsisted of 500 mM Tris, pH 7.5, 1.5 M NaCl and 0.2% Tween-20. BlockerA was made up of bovine serum albumin in Tris Wash Buffer. Read Buffer Twas used according to manufacturer's instructions. The Whole Cell LysateKits used were phospho-Akt Ser473 (K11100D. Lot K0011749) and totalERK1/2 (K11107D, Lot K0011698) as a loading control.

Equipment

12-well multichannel pipettes (30-300 μl) from Eppendorf were used.Assay plates were agitated on a TiMix 5 control (Edmund Buhler).Electrochemiluminescence detection was performed on a SECTOR Imager 6000(Meso Scale Discovery).

Measurement of Phospho-Akt Ser473 Experimental Procedures

Protein assay: protein concentration was determined using the BCA(bicinchoninic acid) Protein Assay kit according to the manufacturer'sinstructions, Briefly, duplicate 10 μl samples of bovine serum albumin(BSA) standards, blank and lysates were incubated in a 96-well platewith 0.2 ml working reagent for 30 min at 37° C. Plates cooled to roomtemperature for 5 min and absorbance at 562 nm measured in a multi-modeplate reader. Protein concentrations were calculated using FlexStation 3software (SoftMax Pro version 5.3). Protein concentration of lysates wasdetermined from a standard curve (BSA) using a linear curve fit.

Phosphoprotein assay: protein phosphorylation was determined using theMULTI-SPOT® Assay System (Meso Scale Discovery), providing simultaneousdetection of phosphorylated and total proteins. Briefly, captureantibodies against phosphorylated and total protein are patterned ondistinct spots in the same well of 96-well plates. Sandwich immunoassayand electrochemiluminescence detection technology are combined tomeasure intensity of the emitted light from phosphorylated and totalprotein spots. For analysis of phosphor-Akt Ser473 was performedaccording to the manufacturer's instructions. The optimal amount ofprotein was determined at 5 μg lysate per well for ERK1/2 and 10 μg/wellfor phospho-Akt Ser473. Plates were blocked with 25 μl/well Blocker Afor 1 h at room temperature with gentle agitation. During this time, thelysates were thawed and diluted to the desired protein concentration incomplete Tris Lysis Buffer. Plates were washed four times in Tris WashBuffer and 25 μl lysate per well added. Plates were incubated for 1-3 hat room temperature with agitation according to the manufacturer'srecommendations. Plates were washed four times with Tris Wash Buffer,followed by addition of 25 μl/well of the respective detection antibodyand incubation for 1 h at room temperature, with agitation. After afinal four washes with Tris Wash Buffer 150 μl/well, Read Buffer T wasadded, and plates read on a SECTOR Imager 6000 plate reader.

Assessment of Effects of Phospho-Akt Ser473

The mean background signal from each plate was subtracted from averagedraw data. The amount of total protein phosphorylaled was expressed as %phosphoprotein according to the manufacturer's (Meso Scale Discovery)Instructions.

Results

Levels of phospho-Akt Ser473 were determined in IgE sensitized andantigen stimulated cells after treatment without (positive control) orwith 1, 5 and 25 μM test compound and normalized to levels of total Akt.Concentration-dependent inhibition of Akt phosphorylation on Ser473 wasdemonstrated, as shown in FIG. 2. Table 2 shows levels of normalizedphospho-Akt Ser473 as a percentage of those in the positive control.

TABLE 2 Inhibition of Akt phosphorylation on Ser473 by compounds 1a and1c Level of phospho-Akt Ser473 (% positive control) Compound 1 μM 5 μM25 μM 1a 103.1 ± 17.4 57.0 ± 15.2 13.2 ± 4.2 1c  83.2 ± 17.3 11.7 ± 7.7  2.2 ± 1.4

Percentage of total Akt phosphorylated on Ser473 expressed as percentageof control untreated cells, after Induction with IgE and antigen for 15min.

A dose-dependent decrease in levels of phospho-Akt Ser473 was observedafter treatment with all compounds outlined In Table 2. Thus, thecompounds according to the invention can be used to reduce levels ofactivated Akt and, accordingly, are useful in the medical interventionin indications In which hyperactivated Akt plays a pathogenic role, suchas inflammatory and allergic diseases.

Example 29: Inhibition of the Delayed-Type Hypersensitivity (DTH)Reaction in Mice Introduction

The anti-inflammatory and anti-allergic effects of compound 1a wasassessed in a mouse model of skin delayed-type hypersensitivity (DTH)reactions and compared to a vehicle control and to the reference drugdexamethasone. DTH reactions are antigen-specific cell-mediated immuneresponses, driven primarily by T helper type 1 (Th1) cells, similar tothe tuberculin immunization response. The immune reaction induced by anovalbumin challenge to animals previously sensitized with ovalbumin inComplete Freund's Adjuvant, is characterized by swelling (edema) at thesite of challenge, e.g. the mouse ear. Dexamethasone, ananti-inflammatory steroid, reduces cell-mediated immune responses andwas employed to validate the responsiveness of the assay topharmacological treatment

Materials and Methods Materials

Ovalbumin (fraction V, lyophilized powder), complete Freud's adjuvant(CFA) and methylcelluiose were obtained from Sigma-Aldrich,dexamethasone from Pharmaceutical Works Polfa (Pabianice, Poland).

Animals

Female BALB/cJW mice were bred at the University of Lodz, Lodz, Polandand housed in groups of 8 in makrolon cages with a 12 h light-darkcycle. Mice were given free access to food (Agropol S. j., Motycz,Poland) and water.

Antigen Sensitization and Challenge

Group size was n=8 mice unless otherwise stated. Test compound wasfreshly prepared before administration.

Sensitization: The protein antigen, ovalbumin, was reconstituted in PBSat 4 mg/ml. An ovalbumin-CFA emulsion was prepared by mixing the proteinsolution with the CFA suspension at a ratio of 1:1, using two Luer-locksyringes. The emulsion was tested by putting a drop of emulsion ontoPBS; if the emulsion remained as a light droplet on the PBS, theemulsion was deemed ready. Mice were sensitized by subcutaneouslyinjecting 25 μL of emuision into each side of the tail (100 μg ovalbuminper mouse).

Challenge: On the sixth day after sensitization, DTH was elicited bychallenging animals subcutaneously (gauge 30 needle, B. Braun Melsungen,Melsungen, Germany) in the left ears with 10 μL of a 1% suspension ofheat-aggregated ovalbumin (HOVA) (100 μg ovalbumin per mouse). The rightears were administered subcutaneously with PBS and served to determinethe individual differences in ear thicknesses. HOVA was prepared byheating a 5% solution of ovalbumin in saline for 1 h at 80° C. withoccasional swirling. After cooling to room temperature andcentrifugation (400 g, 10 min at 4° C.), the pellet was washed twicewith saline, resuspended at 2% in PBS and aliquots stored at −30°C.Before injection, HOVA was diluted with an equal volume of PBS andsonicated. Ear thickness was measured with a precise spring-loadedcaliper (Art. No, 7309, Mltutoyo, Kawasaki, Japan) before challenge, and24 h after challenge.

Sensitization, challenge and ear thickness measurement were performedunder anesthesia (ketamine 80 mg/kg plus xylazine 8 mg/kg,intraperitoneal).

Compound Administration

The anti-inflammalory effects of compound 1a were compared to a vehiclecontrol (0.5% methyl cellulose solution) and to the reference drug,dexamethasone. Test compound was given orally by gavage (Art. No.432093, Harvard Apparatus GmbH, March-Hugstetten, Germany) as follows:16 h prior to ovalbumin sensitization, a loading dose of 100 mg/kg wasadministered; the first maintenance dose of 25 mg/kg was given 3 hbefore sensitization (day 0) and on each of the next five consecutivedays (day 1 to 5) as well as on the day of antigen challenge (day 6) (atotal of 8 administrations). Three hours after the last dose, theantigen challenge was performed on the ears as described above.Dexamethasone was given at 1 mg/kg orally by gavage 3 h beforesensitization and once daily with the final dose given 3 h prior toantigen challenge (a total of 7 administrations). All administrationswere given in a volume of 10 mL/kg.

Quantification of Assay Results

To account for individual variability, the increase in right earthickness, before and 24 h after administration of PBS, was subtractedfrom the HOVA-induced increase in left ear thickness. The increase inear thickness was calculated by the difference between ear thicknessbefore and 24 h after antigen challenge. Percent inhibition of earswelling was calculated according to the following formula;

% inhibition=100x (IET_(vehicle)−IET_(compound))/IET_(vehicle)

whereIET=(ET_(24 hrs pc)−ET_(predose))_(HOVA-treated ears)−(ET_(24 hrs pc)−ET_(predose))_(PBS-treated ears)

(IET, increase ear thickness; ET, ear thickness; pc, post challenge)

Statistical evaluation

Mean and standard deviation (SD) were calculated from individual earedema values. Statistical evaluation was a one-way analysis of variance(ANOVA) with Dunnett's post hoc test or Student's t-test whereappropriate.

Results

Suppression of mouse ear swelling by compound 1a and dexamethasone,compared to vehicle control is shown in Figure 3, Table 3 summarizes theinhibition of DTH for compounds 1a.

TABLE 3 Effect of compound 1a on ear swelling in the DTH response inmice. Compound Inhibition of mouse ear swelling 1a, 100 mg/kg 32*Dexamethasone, 1.0 mg/kg 49* *p < 0.01 vs. vehicle control (Dunett'spost hoc test)

Dexamethasone administered orally at a dose of 1 mg/kg, once daily overthe whole sensitization period resulted in a significantly reduced DTHresponse, with inhibition of 49%. Such high dosing (overdose) is,however, not suitable for treatment of humans due to severe side effectsof the corticosteroid and was only used to validate the responsivenessof the model. In addition, in the course of the current study,administration of dexamethasone resulted in a significant loss in bodyweight of 4.4% (p<0.01 vs. vehicle control with the paired Student'st-test), a typical sign of corticosteroid toxicity. In contrast, notoxic side-effects of compound 1a were observed during the course of thestudy. Compound 1a, administered orally twice daily over the wholesensitization period at 20 mg/kg (loading dose 100 mg/kg), significantlyreduced the DTH response by 32%. Hence, compound 1a was able to producean inhibition equivalent to 65% that of a high dose of dexamethasone.

The reduction of DTH response obtained by treatment with compound 1ademonstrates that the compounds according to the invention and, inparticular compound 1a, are effective in the pharmaceutical interventionin allergic and inflammatory diseases involving antigen-specificcell-mediated immune responses.

Example 30: Inhibition of the Allergic Contact Dermatitis InflammatoryResponse in Mice Introduction

The anti-inflammatory and anti-allergic effects of compound 1a wereassessed in a mouse model of allergic contact dermatitis, a responsedriven primarily by T helper type 2 (Th2) cells. It has beendemonstrated that BALB/c mice are susceptible to the allergentoluene-2,4-diisocyanate (TDI), producing an inflammatory condition ofthe skin with similar aspects to that of human atopic dermatitis (Baumeret al., J Pharm Pharmacol, 55:1107-1114 (2003); Baumer et al., Br JDermatol. 151:823-830 (2004); Ehinger et al., Eur J Pharmacol. 392:93-99(2000)). In this model, an allergic dermatitis response is obtained bysensitizing mice to TDI and subsequently challenging them with antigenby topical administration onto the ears. A quantitative assessment ofanti-inflammatory and anti-allergic effects of topically or orallyadministered test compounds is possible by measuring the resulting earswelling, i The advantages of the allergic contact dermatitis model(Zdliner et al., Bioessays 26:693-6 (2004)) are reproducibility andreliability (>90 % of BALB/c mice respond to sensitization), a shortinduction protocol, quantitative assessment by measuring ear thickness,atopic dermatitis-like skin lesions can be induced, and clinicallyrelevant pharmaceuticals, such as corticosteroids,calcineurin-inhibitors and PDE4-inhibitors, are effective in this model.

Materials and Methods Animals

Female BALB/c-mice were obtained from Charles River (Sulzfeld, Germany)at age 8 weeks. All animals were housed in groups of eight per cage at22° C. with a 12 h light/dark-cycle. Water and a standard diet(Aitromin, Lage/Lippe, Germany) were available ad libitum. All animalswere acclimatized for one week before experimental procedures werecommenced.

TDI Sensitization, Allergen Challenge and Mouse Ear Swelling Test

Experimental procedures for BALB/c mice housing, TDi sensitization andchallenge, and measurement of ear thickness were performed as previouslydescribed (Baurner et al., J Pharm Pharmacol. 55:1107-1114 (2003)) withthe following modifications. For active sensitization, 100 μL of 5%(w/v) TDI was administered to the shaved and stripped abdominalepidermis on day one, and for the next three consecutive days, 50 μL of5% (w/v) TDI was applied. The allergic reaction was boosted 21 dayslater by application of 50 μL of 0.5% (w/v) TDI. For the examination oftest compound effects, the left ears were used for the TDI challenge (20μL of 0.5% in acetone) and ear thickness measured 3 h before and 24 hafter challenge.

Compound Administration for Systemic Treatment

Group size was n=7 mice unless otherwise stated. Test compounds werefreshly prepared before administration.

Administration time: to determine optimal time for administrationtreatment groups were treated orally by gavage with 100 mg/kg ofcompound 1a (suspended in phosphate-buffered saline (PBS), 10 mL/kg) 4or 16 h before topical TDI challenge. Vehicle treated mice received PBS(10 mL/kg) orally, 4 h before challenge.

Dose-response: two groups of mice were treated orally with compound 1aat 20 mg/kg or 100 mg/kg suspended in PBS, 4 h before topical TDIchallenge. Vehicle treated mice received PBS orally 4 h beforechallenge.

Compound Administration for Topical Treatment

Compound 1a was administered to two groups of mice topically in 20 μl ofa 2 % or 6% solution in acetone/water (1:1). The solution was applied, 2h before topical TDI challenge by administration of 10 μl onto each ofthe inner and outer surfaces of the left ears. A vehicle group (n=7) wastreated with acetone/water (1:1).

Determination of Local Lymph Node Weight and Cell Count

Directly after sacrifice, the ear draining lymph node (Ln. auricufaris)was prepared and excised. Organ weight was determined by means of ananalytical balance (Kern, Balingen, Germany). Single cell suspensionswere prepared by means of a glass potter (VWR, Darmstadt, Germany) andcells were counted with a hemocytometer (Neubauer, VWR, Germany).

Statistical Evaluation

Mean and standard error of the mean (SEM) were calculated fromindividual ear edema values. Statistical evaluation was a one-wayanalysis of variance (ANOVA) (if the test for normal distribution waspassed) or the Kruskal-Wallis one-way ANOVA on Ranks (if the normaldistribution test failed), Both were followed by a post-hoc test(Dunnett's method or Dunn's test, respectively). A p<0.05 was consideredto be significant.

Results

Suppression of mouse ear swelling by compound 1a after oraladministration, compared to vehicle control is shown in FIG. 4A. Table 4summarizes inhibition of the allergic contact dermatitis response bycompound 1a.

TABLE 4 Effect of orally administered compound 1a or ear swelling in theallergic contact dermatitis response in mice. Compound Inhibition ofmouse ear swelling Administration time (oral) 1a, 100 mg/kg, 4 h 51.7*1a, 100 mg/kg, 16 h 32.2 *p < 0.05 vs. vehicle control (Dunnett's posthoc test) compared to vehicle

In the administration time study with oral administration, compound 1areduced ear swelling significantly (52% of vehicle control) whenadministered 4 h before challenge, as also shown in FIG. 4A.

Compound 1a had a significant impact on the TDI induced inflammatoryreaction in a pilot study at 100 mg/kg. Thus, the compounds according tothe invention and, in particular compound 1a, are particularly effectiveand thus useful for the oral pharmaceutical intervention in inflammatorydiseases, in particular in atopic dermatitis. Suppression of mouse earswelling by compound 1a after topical administration, compared tovehicle control is shown in FIG. 4B, Table 5 summarizes inhibition ofthe allergic contact dermatitis response by compound 1a.

TABLE 5 Effect of topically administered compound 1a on ear swelling inthe allergic contact dermatitis response in mice. Compound Inhibition ofmouse ear swelling 1a, 2% 72.0*** 1a, 6% 86.0*** ***p < 0.001 vs.vehicle control (Dunnett's post hoc test) compared to vehicle

Compound 1a topically administered as a solution at 2% or 6% reduced earswelling highly significantly by 72 or 86%, respectively.

One of the most undesirable side-effects of corticosteroidadministration is immunosuppression, which leads to the inability toeffectively address parasitic infection, wound healing and tumor growth.In the current study, the local lymph node reaction after TDI challenge(lymph node weight and cell number) was determined to assess theresponse of immune organs. Systemic treatment with compound 1a at 100mg/kg or topical treatment at 2% or 6% did not have any impact on thelocal lymph node reaction.

In view of the strong effect shown in the allergic contact dermatitismodel, the compounds of the present invention and, including compound1a, are particularly effective and thus useful for the topicalpharmaceutical intervention in inflammatory diseases, in particular inatopic dermatitis.

1-48. (canceled)
 49. A compound of the following formula 1

wherein: R¹ is a C₁₀₋₂₀ hydrocarbon group; R² is a C₁₋₄ alkyl group, andR³ is —H, or R is absent; R⁴ is a C₁₋₆ alkylene group; R⁵ is —SO₃ ⁻,—SO₃H, —PO₃H.sup.—, —PO₃H⁻, —PO₃ ²⁻, —PO₃H₂, —PO₂(OC₁₋₃ alkyl)—,—PO₂H(OC₁₋₃ alkyl), —PO(OC₁₋₃ alkyl)₂, —CO₂ ⁻, —CO₂H or —CO₂(C₁₋₃alkyl); and X is N⁺ or, if R³ is absent, X is N.
 50. The compound ofclaim 49, wherein R⁵ is —SO₃ ⁻ or —SO₃H.
 51. The compound of claim 49,wherein R⁵ is —PO₃ ²⁻, PO₃H⁻, or —PO₃H₂.
 52. The compound of claim 49,wherein R₄ is —(CH₂)₃—.
 53. The compound of claim 49, wherein R₁ is alinear C₁₀₋₂₀ alkyl group, a linear C₁₀₋₂₀ alkenyl group, or a linearC₁₀₋₂₀ alkynyl group.
 54. The compound of claim 49, wherein R¹ is—(CH₂)₁₁—CH₃, —(CH₂)₁₃—CH₃, or —(CH₂)₁₅—CH₃.
 55. The compound of claim49, wherein R² is methyl, and R³ is —H or R³ is absent.
 56. The compoundof claim 49, wherein the compound is a compound of any of the followingformulae:

or a pharmaceutical acceptable salt, solvate or prodrug thereof.
 57. Apharmaceutical composition comprising the compound of claim 49 and apharmaceutically acceptable excipient.
 58. A compound of the followingformula 1

wherein: R¹ is a C₁₀₋₁₁ hydrocarbon group, a C₁₃ hydrocarbon group, aC₁₅ hydrocarbon group, or a C₁₇₋₂₀ hydrocarbon group; R² is a C₁₋₄ alkylgroup, and R³ is —H, a C₁₋₄ alkyl group or R³ is absent; R⁴ is a C₁₋₆alkylene group; R⁵ is —SO₃ ^(−, —SO) ₃H, —PO₃H.sup—, —PO₃ ²⁻, —PO₂ ⁻,—PO₃H₂, —PO₂(OC₁₋₃ alkyl)—, —PO₂H(OC₁₋₃ alkyl), —PO(OC₁₋₃ alkyl)₂, —CO₂⁻, —CO₂H or —CO₂(C₁₋₃ alkyl); and X is N⁺ or, if R³ is absent, X is N.59. The compound of claim 58, wherein R⁵ is —SO₃ ^(− or —SO) ₃H.
 60. Thecompound of claim 58, wherein R¹ is a linear C₁₀₋₁₁ hydrocarbon group, alinear C₁₃ hydrocarbon group, a linear C₁₅ hydrocarbon group, or alinear C₁₇₋₂₀ hydrocarbon group.
 61. The compound of claim 58, whereinR² and R³ are mutually linked to form a piperidine ring together withthe nitrogen atom X to which they are attached, wherein the piperidinering is optionally substituted with —OH, —O(C₁₋₃ alkyl), —O—C(O)—(C₁₋₃alkyl), C₁₋₃ alkyl, —C(O)—NH₂, —C(O)—NH(C₁₋₃ alkyl), —C(O)—N(C₁₋₃alkyl)(C₁₋₃ alkyl), —NH₂, —NH(C₁₋₃ alkyl), or —N(C₁₋₃ alkyl)(C₁₋₃alkyl).
 62. The compound of claim 58, wherein the compound is a compoundof the following formula:

or a pharmaceutically acceptable salt, solvate or prodrug thereof.
 63. Apharmaceutical composition comprising the compound of claim 58 and apharmaceutically acceptable excipient.
 64. A compound of the followingformula 1

wherein: R¹ is a C₁₀₋₂₀ hydrocarbon group; R² is a C₁₋₄ alkyl group, andR³ is —H, a C₁₋₄ alkyl group or R³ is absent; R⁴ is a C₁₋₂ alkylenegroup or a C₄₋₆ alkylene group; R⁵ is —SO₃ ⁻, —SO₃H, —PO₃H.sup—, —PO₃H⁻,—PO₃ ²⁻, —PO₃H₂, —PO₂(OC₁₋₃ alkyl)⁻, —PO₂H(OC₁₋₃ alkyl), —PO(OC₁₋₃alkyl)₂, —C₂ ⁻, —CO₂H or —CO₂(C₁₋₃ alkyl); and X is N⁺ or, if R³ isabsent, X is N. 65.The compound of claim 64, wherein R⁵ is —SO₃ ⁻ or—SO₃H.
 66. The compound of claim 64, R⁵ is —P₃ ²⁻, —PO₃H⁻, or —PO₃H₂.67. The compound of claim 64, wherein R¹ is a linear C₁₀₋₂₀ alkyl group,a linear C₁₀₋₂₀ alkenyl group, or a linear C₁₀₋₂₀ alkynyl group.
 68. Thecompound of claim 64, wherein the compound is a compound of thefollowing formulae:

or a pharmaceutically acceptable salt, solvate or prodrug thereof.
 69. Apharmaceutical composition comprising the compound of claim 64 and apharmaceutically acceptable excipient.